Centrifuge system for separating suspended cells

Abstract

An apparatus for separating a cell suspension into a centrifuge liquid and a concentrate includes a disposable structure (178, 240, 250, 370, 414) releasably positioned in a cavity within a solid-walled, rotatable centrifuge bowl (172). The bowl and a portion of the disposable structure rotate about an axis (174, 428). A stationary inlet supply line (184, 430), a centrifuge discharge line (212, 436), and a concentrate discharge line (230, 448) extend along the axis of the rotating disposable structure. A centrifuge pump (208, 438) is fluidly connected to the centrifuge discharge line. A concentrate centrifuge pump (216, 450) is fluidly connected to the centrifuge discharge line. At least one centrifuge channel (380, 454) and a centrifuge pump chamber (376, 452) are configured to facilitate the flow of the cell concentrate.

Description

FIELD OF THE INVENTION

[0001] FIELD OF THE INVENTION The present invention relates to the centrifugation of raw materials. An exemplary configuration of the present invention relates to an apparatus for separating cells in suspension by centrifugation. [Background technology]

[0002] Devices and methods for separating cells in suspension are useful in many technical environments. There is room for improvement in such devices and methods. Summary of the Invention

[0003] Exemplary configurations disclosed herein relate to devices and methods for centrifuging cells in large-scale cell culture using pre-sterilized, disposable flow path components. The centrifuge used in the present invention can be any solid-wall centrifuge that uses pre-sterilized, disposable components and is capable of processing cell suspensions at high cell concentrations.

[0004] Exemplary configurations utilize rotatably fixed supply / discharge elements. Disposable elements often have a flexible membrane attached to a rigid frame with a core having an expanded diameter. The disposable element may also include at least one cardiac pump. The disposable structure may be supported within a rigid, multi-use bowl with a frusto-conical interior. These structural elements allow the system to maintain sufficiently high angular velocities to achieve settling velocities that efficiently process highly concentrated cell culture media streams. Feed characteristics, such as minimal turbidity, allow for continuous or semi-continuous cell concentrate discharge, enabling overall production rates greater than currently available. The exemplary structures and methods described herein can be effectively implemented with minimal risk of contamination. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] U.S. Published Application No. 2010 / 0167388 [Patent Document 2] U.S. Patent No. 10,384,216 [Patent Document 3] U.S. Patent No. 9,222,067 [Patent Document 4] U.S. Patent No. 6,615,590 [Patent Document 5] U.S. Patent No. 9,427,748 [Non-patent literature]

[0006] [Non-Patent Document 1] “Microprocessor Architecture, Programming, and Applications with the 8085” by Ramesh S. Gaonker (Prentic Hall, 2002) [Brief explanation of the drawings]

[0007] [Figure 1] FIG. 1 is a schematic diagram illustrating an exemplary configuration of a centrifuge system having disposable multi-use components. [Figure 2] FIG. 2 is a detailed view of the upper flange area of ​​the centrifuge system configuration of FIG. 1, illustrating how the flexible chamber material is sealed to the surface of the flange. [Figure 3] FIG. 3 is an isometric cutaway view of the disposable core and top flange of the centrifuge system configuration shown in FIG. [Figure 4] FIG. 4 is a schematic diagram illustrating the configuration shown in FIG. 1 in which the pumping chamber of the centrifuge system has accelerator fins. [Figure 5] FIG. 5 is an isometric view of the top of the pump chamber in the centrifuge system configuration shown in FIG. [Figure 6] FIG. 6 is an isometric cutaway view of the core and upper and lower flanges of a disposable centrifuge system with an expanded core diameter (for shallow pool centrifugation). [Figure 7]FIG. 7 is an isometric view of the delivery accelerator of FIG. [Figure 8] FIG. 8 is an isometric cutaway view of the core and upper and lower flanges of a disposable centrifuge system with a standard core diameter, a feed accelerator with curved blades, and an oval bowl. [Figure 9] FIG. 9 is an isometric view of the delivery accelerator of FIG. [Figure 10] FIG. 10 is a schematic diagram showing a portion of a centrifuge system with continuous concentrate discharge. [Figure 11] FIG. 11 is a schematic diagram showing another configuration having a centrifuge system with continuous discharge of concentrate. [Figure 12] FIG. 12 is a schematic diagram showing a centrifuge system with a diluent introduction and continuous concentrate discharge. [Figure 13] FIG. 13 is a schematic diagram showing yet another configuration of a centrifuge system with continuous concentrate discharge and a centripetal pump throttling mechanism. [Figure 14] FIG. 14 is an isometric cutaway view of the core and top flange of a disposable centrifuge system with a core and a feed accelerator with straight blades. [Figure 15] FIG. 15 is an isometric view of the delivery accelerator of FIG. [Figure 16] FIG. 16 is an isometric cutaway view of a centrifuge system with continuous discharge of another concentrate. [Figure 17] FIG. 17 is an isometric exploded view of another centripetal pump. [Figure 18] FIG. 18 is an isometric view of another centripetal pump plate with a spiral flow path therein. [Figure 19] FIG. 19 is a schematic diagram illustrating a centrifuge system that operates to ensure positive pressure is maintained within the centrifuge core cavity. [Figure 20] FIG. 20 is a schematic diagram illustrating a simplified exemplary logic flow performed by at least one control circuit shown in FIG. [Figure 21] FIG. 21 is a cross-sectional schematic diagram showing another centrifuge system with continuous centrate and concentrate discharge. [Figure 22] FIG. 22 is a cross-sectional schematic diagram showing yet another centrifuge system with continuous centrate and concentrate discharge. [Figure 23] FIG. 23 is a cross-sectional schematic diagram showing yet another centrifuge system with continuous centrate and concentrate discharge. [Figure 24] FIG. 24 is a schematic diagram illustrating an exemplary continuous permeate and concentrate discharge centrifuge system. [Figure 25] FIG. 25 is a schematic diagram illustrating the logic flow corresponding to the exemplary control system of FIG. [Figure 26] FIG. 26 is a cross-sectional view of an exemplary top portion of a disposable centrifuge design having a concentrate / fraction dam within the separation chamber. [Figure 27] FIG. 27 is a cross-sectional view showing an exemplary top portion of a disposable structure with vanes in the centrate and concentrate pumping chambers to control the radial position of the air / liquid interface. [Figure 28] FIG. 28 is a perspective view of an exemplary concentrate or fractionate pump chamber surface having multiple chamber vanes. [Figure 29] FIG. 29 is an axial cross-sectional view of an exemplary upper portion of a disposable structure similar to that shown in FIG. 27 showing the location of the air / liquid interface. [Figure 30] FIG. 30 is an axial cross-sectional view of an exemplary upper portion of a disposable structure including an air flow channel that maintains pressurized air within an air pocket. [Figure 31] FIG. 31 is a schematic diagram illustrating an exemplary centrifuge system with backpressure control of centrate flow. [Figure 32] FIG. 32 is a schematic axial cross-sectional view of yet another continuous centrate / concentrate discharge centrifuge system. [Figure 33] FIG. 33 is a cross-sectional schematic view showing the top of the system shown in FIG. [Figure 34] FIG. 34 is a cross-sectional schematic view similar to FIG. 32, but showing the system in operation and with the annular cell concentrate region established within the separation chamber. [Figure 35] FIG. 35 is an exterior front perspective view of yet another disposable centrifuge structure. [Figure 36] FIG. 36 is a cross-sectional view of the disposable structure shown in FIG. [Figure 37] FIG. 37 is an exploded view showing the upper disk portion of the disposable structure shown in FIG. [Figure 38] FIG. 38 is a perspective view of the underside of the upper disk-shaped portion of the structure shown in FIG. DETAILED DESCRIPTION OF THE INVENTION

[0008] In the field of cell culture as applied to biopharmaceutical processes, cells must be separated from a fluid medium, such as the fluid in which the cells are cultured. The desired product of cell culture can be a molecular species excreted by the cells into the medium, a molecular species that remains within the cell, or the cells themselves. At the production scale, the initial stages of the cell culture process typically occur in a bioreactor, which can operate in either batch or continuous mode. Repeated batch processes are also possible. The desired product often must be separated from other process components prior to final purification and product formulation. Cell harvest is a general term referring to these cell separations from other process components. Additionally, clarification refers to cell separations where the goal is a cell-free supernatant (or centrifuge), and cell recovery refers to separations where the goal is a cell concentrate. Exemplary configurations disclosed herein relate to cell harvesting in large-scale cell culture systems.

[0009] Cell harvesting and separation methods include batch centrifugation, intermittent centrifugation, continuous centrifugation, semi-continuous centrifugation, tangential flow filtration (TFF), and depth filtration. Centrifuges for cell harvesting in large-volume production-scale cell culture have historically been complex, multi-use systems requiring clean-in-place (CIP) or sterilize-in-place (SIP) techniques to establish a sterile environment to prevent microbial contamination. Smaller systems are available for laboratory-scale and continuous cell harvesting processes. The UniFuge centrifuge system manufactured by Pneumatic Scale Corporation and described in U.S. Published Application No. 2010 / 0167388 (incorporated herein in its entirety) successfully performs cell harvesting in batch cultures at rates ranging from 3 to 30 liters per minute using intermittent processing for volumes up to approximately 2000 liters. U.S. Patent Nos. 10,384,216 and 9,222,067 (both owned by Pneumatic Scale Corporation, the assignee of this application) are also incorporated herein in their entirety. Generally, intermittent processing requires periodic cessation of both the centrifuge bowl rotation and the feed flow to allow the concentrate to drain. This processing method is usually successful for low-concentration, high-viability cultures. It allows for the processing of large batches, and the cell concentrate can be drained relatively quickly and completely.

[0010] It is sometimes necessary to harvest cells from high-concentration and / or low-viability cell cultures that contain high concentrations of cells and cell debris in the feed, sometimes referred to as "high-turbidity feed." Such high-turbidity feed can be slow to process in some centrifuge systems for the following reasons: (1) The feed flow rate must be slowed down to increase the residence time in the centrifuge to separate small cell debris particles. (2) As the concentration of both cells and cell debris increases, the cell concentrate fills the bowl faster, requiring the bowl to be stopped and the concentrate to be drained. These factors combine to reduce net throughput and potentially lead to unacceptably long cell harvest processing times, which in turn increases costs, increases centrifugation times, increases the risk of product contamination, and can result in loss of low-viability cell cultures.

[0011] High cell and cell debris concentrations in the feedstock result in very high viscosity cell concentrates. This makes it difficult to completely drain the cell concentrate from the centrifuge, even with extended drain cycles. In some cases, extended buffer rinse cycles are sufficient to completely drain the concentrate, but either or both of these adjustments to the drain cycles may be necessary, lengthening processing times. This increases the complexity and cost of large-volume cell culture processing.

[0012] Scaling up system size by increasing bowl size and extending the feed portion of the intermittent processing cycle is often impractical because the cell concentrate discharge cycle is correspondingly longer. Another limitation arises that prevents simple geometrical scale-up and variability in scaling up appropriate fluid dynamic factors. For any centrifuge, the maximum processing speed depends on the settling velocity of the particles to be separated. This settling velocity is given by a variation of Stokes' law, defined by Equation 1: Formula 1 TIFF0007813713000001.tif12150 where v is the settling velocity, Δρ is the solid-liquid density difference, d is the particle diameter, r is the particle's radial position, ω is the angular velocity, and μ is the liquid velocity. For scale-up geometries, changing the bowl radius changes the maximum radial position r that a particle can occupy. Thus, if the other parameters in Equation 1 are held constant, increasing the bowl radius increases the average settling velocity and increases throughput for a given separation efficiency. However, as the radius increases, the angular velocity of the bowl becomes more difficult to maintain due to the higher feed strength that may be required and other engineering limitations. Both the average settling velocity and the increased throughput (proportional to radius) decrease when the decrease in angular velocity exceeds the square root of the proportionally increasing radius.

[0013] One engineering limitation that needs to be considered is that the angular velocities required to rotate a larger bowl are difficult to achieve in practice, as they require a larger and more costly centrifuge drive platform.

[0014] Furthermore, if the angular velocity is kept constant while the radius increases, the force propelling the cells toward the centrifuge wall also increases. Rotating the bowl at a sufficiently high angular velocity to achieve the desired processing efficiency increases the stress on the vessel walls and the cells within the vessel. For cells, excessively high cell packing can result in cell damage. Cell damage can be detrimental in applications where cell viability must be maintained and can lead to product contamination in solution during separation. High viscosity resulting from excessively high cell concentrations can also be detrimental to the complete evacuation of the cell concentrate.

[0015] Exemplary configurations include devices and methods for continuous or semi-continuous centrifugation of low-viability cell suspension cultures containing high concentrations of cells and cell debris at speeds suitable for processing large volumes of cell suspensions on an industrial scale. Some exemplary centrifuges are pre-sterilized, disposable designs and can process such cell suspensions at flow rates exceeding 20 liters per minute. This flow capacity allows for total run times of 2-3 hours for a 2000-liter bioreactor. Exemplary configurations of disposable centrifuge systems can process approximately 300-2,000 liters of fluid while operating at approximately 20-40 liters per minute.

[0016] 1 shows a disposable centrifuge structure 1000. The centrifuge structure 1000 includes a core structure 1500 (best seen in FIG. 3) having a core portion 1510, an upper flange 1300, a lower flange 1200, and a flexible liner 1100 sealed to both the upper flange 1300 and the lower flange 1200. The centrifuge structure 1000 includes a centripetal pump 1400 having a pair of stationary paring discs 1410 within a rotary pump chamber 1420, and a rotary mechanical seal 1700.

[0017] The centrifuge structure 1000 also includes a supply / drain system 2000. This system 2000 includes a number of coaxial pipes centered around the axis of rotation 1525 (shown in FIG. 12) of the centrifuge 1000. The innermost portion of the supply / drain system 2000 includes a supply pipe 2100. A number of secondary pipes coaxially surround the supply pipe 2100 and may include pipes or fluid lines for a centrate drain 2200, a concentrate drain 2500 (see, e.g., FIG. 12), or a diluent supply 5000 (see, e.g., FIG. 12). Each portion of the supply / drain connection may be fluidly connected to a portion of the interior of the centrifuge 1000 by appropriate fluid connections and may further be fluidly connected to the coaxial pipes to remove or add a centrate, concentrate, or diluent from the system.

[0018] The upper and lower flanges 1300, 1200 shown in FIG. 1 have conical bowls that are axially aligned with and concave relative to the core 1510. The core 1510 has a generally cylindrical body with a hollow cylindrical center sized to receive the supply line 2100 having an axis 1525 (best seen in FIG. 12). The upper flange 1300, core 1510, and lower flange 1200 can be of unitary construction, providing a more rigid support for the flexible liner 1100, sometimes referred to as a membrane. In other configurations, the core 1500 can be comprised of multiple components. In other configurations, the core 1510 and upper flange 1300 can be comprised of a single component, with the lower flange 1200 being a separate component. Alternatively, the core 1510 and lower flange 1200 can be comprised of a single component, with the upper flange 1300 being a separate component.

[0019] An exemplary unitary core 1510 / top flange 1300 configuration is shown in Figure 3. This unitary member can be joined to the bottom flange 1200 to form the internal support structure 1500 of the disposable member of the centrifuge 1000. This structure secures the flexible liner 1100 at its top and bottom around the rigid or semi-rigid fixed internal support structure 1500. The exterior of the flexible liner 1100 is also supported by the bowl and cover walls of the multi-use structure 3000 when the centrifuge system is in use.

[0020] The exemplary separation chamber 1550 is a generally cylindrical open chamber that is generally bounded by the outer surface 1515 of the core 1510 and the flexible liner 1100, as well as by the upper surface 1210 of the lower flange 1200 and the lower surface 1310 of the upper flange 1300. The separation chamber 1550 is fluidly connected to the supply piping 2100 by a hole 1530 that extends from the central cavity 1520 of the core 1510 to the outer surface 1515 of the core 1510. The separation chamber 1550 is also fluidly connected through the core 1500 to the pump chamber 1420 by a similar hole 1540. In this embodiment, the hole 1540 is angled upward toward the pump chamber 1420 and opens into the separation chamber 1550 directly below the junction of the core 1510 and the upper flange 1300. The holes 1420 or 4420 enter the pump chamber at an angle other than an upward angle, such as horizontally or at a downward angle, as shown in Figure 12. Additionally, in some configurations, the holes 1420, 4420 may be replaced by slits or gaps between the accelerator fins.

[0021] FIG. 1 also shows the supply / drain device 2000 with a supply tubing carrier 2300 through which the supply tubing 2100 extends to a position shown in FIG. 3 near the bottom of the centrifuge 1000. In this position, the supply tubing 2100 can perform both the supply and drain functions without moving. Shear forces during the feeding process can be minimized by careful design of the gap between the nozzle 2110 of the supply tubing 2100 and the upper surface 1210 of the lower flange 1200, as well as the diameter of the nozzle 2110 of the supply tubing 2100 and the angular speed of the centrifuge. U.S. Pat. No. 6,615,590 (incorporated herein by reference) discloses how to select the appropriate relationship to minimize shear forces. Other suitable supply tubing that minimizes shear forces generated in response to the feeding of liquid cell culture media into a rotary centrifuge, known to those skilled in the art, can also be used.

[0022] Also shown in Figure 1 is a centripetal pump 1400 that discharges the centrate via a centrate discharge channel 2200. In the configuration shown in Figure 1, the centrate pump 1400 is located on the upper flange 1300 within a pump chamber 1420. The pump chamber 1420 is defined by the upper surface 1505 of the core 1510 and the inner surfaces 1605, 1620 of the centrifuge cover 1600. The centrifuge cover 1600 can include a cylindrical wall portion (shown in Figure 5) and an engaging cap portion 1610 that generally takes the form of a circular disk (shown in Figure 5). The centrifuge cover 1600 can be formed as a single unit or can be formed from separate components.

[0023] As will be described in more detail below, other configurations may vary in the shape and location of the separator pump chamber 1420. The pump chamber 1420 is an axially symmetric pump chamber near the top of the core structure 1500, which is fluidly connected to the separation chamber 1550 through holes or slits 1530 that extend into the separator pump chamber 1420 from near the exterior of the core 1515. In some configurations, the separator pump chamber 1420 may be recessed within the separation chamber 1550, as best seen in Figures 11 and 12.

[0024] The exemplary separator pump 1400 includes a pair of pairing discs 1410. The pairing discs 1410 are two thin, circular discs (plates) axially aligned with the axis 1525 of the core structure 1500. In the configuration shown in FIGS. 1-5, the pairing discs 1410 are held stationary relative to the centrifuge structure 1000 and separated from each other by a gap 1415 (1415 in FIG. 15). This gap 1415 between the pairing discs 1410 fluidly connects and removes separator from the centrifuge 1000, where it flows between the pairing discs 1410 and into a hollow, cylindrical separator discharge channel 2200 surrounding the supply tubing carrier 2300, which terminates in a separator outlet 2400.

[0025] The exemplary disposable centrifuge structure 1000 is housed within a multi-use centrifuge structure 3000. This structure 3000 includes a bowl 3100 and a cover 3200. The walls of the centrifuge bowl 3100 support the flexible liner 1100 of the centrifuge 1000 during rotation of the centrifuge 1000. To achieve this, the exterior structure of the disposable centrifuge 1000 and the interior structure of the multi-use structure 3000 mate with each other. Similarly, the top surface of the upper flange 1200, the exterior of the upper portion of the core 1510, and the lower portion of the wall 1640 of the centrifuge cover 1600 mate with the interior surface of the multi-use bowl cover 3200, which provides support during rotation. As explained in more detail below, the presence of the multi-use bowl 1300 and bowl cover 3200 prevents shear forces from ripping through the liner 1100 and severing its connection with the disposable centrifuge 1000. In some configurations, an existing multi-use structure 3000 can be integrated into the disposable process by selecting a matching disposable centrifuge 1000. In other configurations, the multi-use structure 3000 may be specifically designed for use with the disposable centrifuge 1000.

[0026] FIG. 2 shows portions of an exemplary structure of the upper flange 1300, plastic liner 1100, and cover 3200 of the versatile centrifuge structure 3000 to illustrate the sealing of the flexible liner 1100 to the upper flange 1300. The flexible liner 1100 can be made of a thermoplastic elastomer such as polyurethane (TPU) or other tough, tear-resistant, biocompatible polymer, while the upper and lower flanges 1300, 1200 can be made of a rigid polymer such as polyetherimide, polycarbonate, or polysulfone. The flexible liner 1100 is a thin sleeve or envelope that extends between and seals to the upper and lower flanges 1300, 1200, forming the outer wall of the separation chamber 1550. The materials used for the liner 1100, upper and lower flanges 1300, 1200, and core 1510 are merely exemplary. Those skilled in the art will be able to select suitable materials with similar properties to known materials.

[0027] A thermal bond attachment process can be used to bond dissimilar materials within the area shown in Figure 2. The flange material is preheated, an elastomeric polymer is placed on top of the heated polymer, and heat and pressure are applied to the elastomeric film liner 1100 at a temperature above the softening point of the elastomeric film liner 1100 to form a thermal bond 1110. In a similar manner, a plastic liner 1100 is bonded to the bottom flange 1200. While a thermal bond 1110 has been described, this is for illustrative purposes only. Other means of forming a similarly strong, relatively permanent bond between the flexible film and the flange material can be substituted, such as temperature, chemicals, adhesives, or other bonding means.

[0028] Exemplary disposable components are pre-sterilized. Thermal seals 1110 maintain the sterility within the disposable chamber while the components are removed from their protective packaging and installed in the centrifuge. During use, flexible, extensible liner 1100 conforms to the walls of reusable bowl 3100. Reusable bowl 3100 provides sufficient support, and flexible liner 1100 provides sufficient resilience, to enable disposable centrifuge 1000 to withstand the high rotational forces generated when the large radius centrifuge 1000 is filled with liquid cell culture media or other suspensions and rotated at high angular velocities to achieve sedimentation rates of approximately 2-40 liters per minute.

[0029] In addition to the thermal seal 1110, sealing ridges or "nubbins" 3210 are present on the bowl cover 3200, compressing the rigid upper flange 1300 with the thermoplastic elastomeric film, further enhancing the seal. The same compression seal is used on the bottom of the bowl 3100, allowing the rigid lower flange 1200 to be sealed with the thermoplastic elastomeric film. These seals support the thermal seal 1110 by isolating it from the shear forces caused by the hydrostatic pressure generated during centrifugation when filling the chamber. The seal between the thermal seal 1110 and the compression nubbins 3210 was tested to 3000 x g, corresponding to a hydrostatic pressure of 97 psi at the bowl wall. The lining is thin and compressible enough that the nubbins 3210 compress and grip the flexible liner 1100, minimizing the risk of tearing near the thermal seal 1110 or the compression nubbins 3210. In one example configuration, the flexible TPU liner has a seal thickness of 0.010 inches and exhibits no tearing or leaking.

[0030] A configuration corresponding to that shown in Figures 1 and 2 was tested in a 5.5 inch diameter bowl. The hydraulic capacity at 2000 x g was >7 liters / minute, and mammalian cells were successfully separated with 99% efficiency at a flow rate of 3 liters / minute.

[0031] In most cases, the upper and lower flanges 1300, 1200 may have a shape similar to that shown in Figure 1, but in some cases, the top surface of a disposable centrifuge may have a different shape, as shown in Figures 10 and 11. In the configuration shown in Figures 10 and 11, both the top flange and bowl cover are disk-shaped, as compared to the generally conical bowl cover 3200, which matches the generally conical top flange 1300. One skilled in the art will be able to apply the sealing techniques of the present invention to sealing surfaces of different shapes.

[0032] FIGS. 4 and 5 illustrate an example configuration with features that improve the efficiency of the centripetal pump 1400. As shown in detail in FIG. 5, the disposable's internal structure, similar to that shown in FIGS. 1 and 2, includes a plurality of radial fins 1630 on the inner surface 1620 of the cap portion 1610 of the pump chamber 1420. The radial fins 1630 may be thin, generally rectangular radial plates that extend perpendicularly from the inner surface 1620 of the cap portion 1610. In the exemplary configuration, six fins 1630 are used, although other configurations may include fewer or more than six fins 1630. In this configuration, the fins form part of the inner surface of the cap 1620, but other configurations may include the upper surface 1620 of the pump chamber 1420, which may take on a shape other than that of the cap 1610. When the centrifuge system is in use, the fins 1630 are positioned within the pump chamber 1420 on the paring disk 1410 of the centripetal pump 1400. These fins 1630 transfer the angular rotation of the centrifuge 1000 to the centrate in the pumping chamber 1420 .

[0033] This results in improved efficiency of the centripetal pump 1400, a more stable gas-liquid interface within the pumping chamber 1420 above the pairing disk 1410, and an increased gas barrier size. This gas barrier is a generally cylindrical gas column that extends outward from the exterior of the supply / drain mechanism 2000, into the pumping chamber 1420, and onto the inner surface of the rotating separation fluid. This increased gas barrier size occurs because the angular velocity of the separation fluid increases, forcing the separation fluid against the centrifuge wall. When the separation fluid rotating within the pumping chamber 1420 comes into contact with the stationary pairing disk 1410, friction can occur, reducing the efficiency of the pump 1400. The addition of multiple radial fins 1630, which rotate at the same angular velocity as the separation fluid, can reduce this speed reduction. This speed reduction results from collisions between the rotating separation fluid and the stationary pairing disk 1410.

[0034] FIG. 6 shows an exemplary configuration of a core structure 1500 for use with highly turbid feedstocks. The core structure 1500 includes a core section 1510, an upper flange 1300, and a lower flange 1200. The core section 1510 includes a cylindrical central cavity 1520 into which the supply tubing 2100 can be inserted. The distance from the central axis 1525 to the outside of the core section 1515 (the core section width, shown by dashed line 6000 in FIG. 6) is longer than the corresponding distance in the configuration shown in FIG. 3. The use of a larger diameter core section 1510 reduces the depth of the separation chamber 1550 (shown by dashed line 6010), allowing the centrifuge 1000 to operate as a shallow pool centrifuge. Generally, the depth 6010 of the separation chamber 1550 is the distance between the outside of the core section 1510 and the flexible liner 1100 as shown in FIGS. 1 and 12. In a shallow pool centrifuge, the depth 6010 of the separation chamber 1550 is small relative to the diameter of the centrifuge. As can be seen from the exemplary configuration shown in FIG. 12, the shallow pool depth 6010 is shallower at the bottom of the separation chamber 1550 and somewhat deeper at the top of the separation chamber 1550 to facilitate removal of the cell concentrate. In some configurations of the present invention, the ratio of the average separation pool depth 6010 to the core width is 1:1 or less. An example of a shallow pool centrifuge is select models of the ViaFuge® centrifuge system manufactured by Pneumatic Scale Corporation. One advantage of a shallow pool centrifuge is that it allows for separation at higher feed flow rates. This advantage is realized by a higher average gravity force for a given bowl inner diameter, which allows for a higher settling velocity at a given angular velocity. The resulting improved separation characteristics are advantageous when separating turbid feedstocks containing high concentrations of cellular debris.

[0035] The embodiment of the core structure 1500 shown in Figure 6 also includes accelerator vanes 1560 as part of the lower flange 1200. Rather than holes 1530 drilled into the solid core 1510 (shown in Figures 10 and 11), the accelerator vanes 1560 (shown in Figure 12) provide another fluid connection between the central cavity 1520 of the core 1510 and the separation chamber 1550.

[0036] In the exemplary configuration of the core structure 1500 shown in FIG. 6, the accelerator vanes 1560 include a plurality of radially spaced, generally rectangular, thin plates 1580 that extend upward from the upper conical surface of the lower flange 1200. The plates 1580 extend upward perpendicular to the base of the core 1510. The plates 1580 also extend generally radially outward from near the axis 1525 of the core 1510. In the exemplary configuration, as best seen in FIG. 7, there are twelve plates 1580. In other configurations, there may be fewer or more than twelve plates. Additionally, in other configurations, the plates 1580 may be curved in the direction of rotation of the centrifuge 1000, as shown in the exemplary configuration of FIG. 9. The inner surface of the lower flange 1200 may be configured to form an oval-shaped accelerator bowl 1590 from which curved plates extend upward. The above configurations are exemplary, and one skilled in the art could combine or modify these configurations in different ways to further benefit from the plates, the shape of the lower flange 1200, and / or the turbidity reduction provided by the buried accelerator.

[0037] Further advantages of the continuous or semi-continuous operation of the disposable centrifuge 1000 are illustrated in FIGS. 10-12. The exemplary configuration shown in FIG. 10 includes a second centripetal pump 4400 for removing a cell concentrate. The centripetal pump 4400 for removing the cell concentrate is positioned above the centripetal pump 1400 for removing the separation solution. The centripetal pump 4400 includes a pump chamber 4420 and a pairing disk 4410. A plurality of holes or continuous slits 4540 extend into the pump chamber 4420 from the upper periphery of the separation chamber 1550, thereby fluidly connecting the upper outer portion of the separation chamber 1550 to the second pump chamber 4420. Like the pump chamber 1400, the pump chamber 4400 may have a different shape than that shown in FIGS. 10-12, but has an overall axially symmetric shape near the upper end of the core structure 1500 that fluidly connects to the separation chamber 1550. Like pump chamber 1400, this pump chamber can be formed in part or entirely by a recess in core structure 1500. If separation fluid pump chamber 1400 is located near the top of core structure 1500, cell concentrate pump chamber 4400 will be located entirely above it. Pump chamber 4400 is fluidly connected to separation chamber 1550 by holes or slits 4540 for removing the cell concentrate. These slits or the like extend from near the outer top wall of separation chamber 1550 to allow recovery of the heavier cell concentrate that is forced thereto by centrifugal force.

[0038] In the configuration shown in FIG. 10, the pairing disk 4410 used in the concentrate discharge pump 4400 has approximately the same radius as the pairing disk 4410 used in the centrate discharge pump 1400 and is rotatably fixed. In configurations such as that shown in FIG. 11, the radius of the pairing disk 4410 in the concentrate discharge pump 4400 is larger than that in the centrate discharge pump 1400, and the pump chamber 4420 is correspondingly larger. Pairing disks with various intermediate diameters can also be used. The optimum diameter depends on the characteristics of the cell concentrate to be discharged. A larger diameter pairing disk provides higher pumping capacity but also generates higher shear forces.

[0039] In the configurations shown in Figures 1, 4, and 10, the paring disk 4410 of the concentrate discharge pump 4400 is rotatably fixed. In other configurations, such as that shown in Figure 11, the paring disk 4410 is rotatable at an angular velocity between zero and the angular velocity of the centrifuge 1000. The desired angular velocity can be controlled by a number of mechanisms well known to those skilled in the art. An example of a control means is an external slip clutch, which allows the paring disk 4410 to rotate at an angular velocity that is a fraction of the angular velocity of the centrifuge 1000. Other means of controlling the angular velocity of the paring disk will be apparent to those skilled in the art.

[0040] In the configurations shown in Figures 1, 4, and 10-12, the gap 1415, 4415 between the pairing disks 1410 and 4410 is fixed. In configurations such as that shown in Figure 13, the gap 1415, 4415 between the pairing disks 1410 and 4410 is adjustable to control the rate at which the permeate and concentrate are removed from the centrifuge 1000. One of each pair of pairing disks 1410 and 4410 is attached to a throttle tube 6100 that can be moved up and down. This throttle tube 6100 can be moved up and down to narrow or widen the gap 1415, 4415 between each pair of pairing disks 1410 and 4410. Additionally, an external peristaltic pump 2510 (not shown) can be attached to the concentrate removal line 2500 (not shown) to facilitate the removal of the concentrate. The pump 2510 can be controlled by a sensor 4430 in the pump chamber 4420. The sensor 4430 (not shown) can be used to control the diluent pump 5150, which allows the withdrawal of concentrate to be synchronized with the addition of diluent.

[0041] FIG. 13 also illustrates a configuration in which the centrate pump 1400 is located at the base of the centrifuge 1000. In the configuration shown in FIG. 13, a centrate reservoir 1555 is formed between the pump chamber 1420 and the flexible liner 1100. Apertures 1530 extend into this centrate reservoir 1555 from the core 1510 below the pump chamber 1420. Furthermore, in the illustrated exemplary configuration, a perforation 1540 extends into the pump chamber 1420 from the separation chamber 1550 adjacent the outer surface 1515 of the core 1510, allowing the separation pump 1400 to remove the centrate. Additionally, a perforation 4540 extends into the pump chamber 4420 between the separation chamber 1550 and its upper outer surface, allowing the cell concentrate to enter the pump chamber 4420 and be removed using the centripetal pump 4400.

[0042] As noted above, in the exemplary configuration shown, the gap 1415, 4415 between the pairing disks 1410, 4410 is adjustable through the use of a throttle tube 6100 connected to one of each pair of pairing disks 4410, 1410. The throttle tube 6100 and one of each pairing disk 4410, 1410 can be moved up and down to narrow or widen the gap 1415, 4415. In the exemplary configuration shown, the throttle tube 6100 is attached to the lower and upper pairing disks of the pairing disk pair 4410, 1410, respectively. In other configurations, this attachment order can be reversed, one centripetal pump can be throttled, or two centripetal pumps can be throttled in parallel (rather than reversed as shown in FIG. 13).

[0043] As can be seen from the configuration shown in Figures 10-12, the walls of solid multi-use bowl 3100 are thicker at the base than at the top, resulting in a frusto-conical interior that can support disposable centrifuge structure 1000 with a smaller radius at the bottom than at the top. Thus, the larger radius at the top of separation chamber 1550 forces the thicker cell concentrate toward the top of separation chamber 1550 and into centripetal pumping chamber 4420. In the illustrated configuration, the frusto-conical shape is achieved by multi-use bowl 3100 having walls that are thicker at the base than at the top. One skilled in the art will recognize that multi-use bowl 3100 with a frusto-conical interior shape can also have walls of uniform thickness, and that multi-use bowl 3100 can be modified to accommodate various interior shapes.

[0044] In the exemplary configurations shown in Figures 10-12, the supply mechanism 2000 also includes an additional flow path for removing cells or cell concentrate. In the configuration shown in Figure 1, a cylindrical flow path 2200 around the supply line 2100 is used to remove the separation liquid. The configurations shown in Figures 10-12 also include a cylindrical coaxial flow path for removing cells or cell concentrate, referred to herein as the cell outlet line 2500. The cell outlet line 2500 surrounds the concentrate outlet flow path 2200. If the centrifuge is designed for use with concentrates that are expected to have very high viscosities, an additional cylindrical coaxial flow path 5000 can be provided around the supply line 2100 to introduce a diluent into the cell concentrate pump chamber 4420 to reduce the viscosity of the concentrate. 12, the diluent channel 5000 comprises a coaxial tube surrounding the cell outlet channel, opening at its lower end into a thin disk-shaped channel 5100 on the pairing disk 4410 and discharging near the outer edge of the pairing disk 4410, thereby providing fluid communication with the pump chamber 4420. Injecting the diluent by this means in this location would restrict the diluent from being introduced into the separation solution, and mixing with and discharging the concentrate, which may be undesirable depending on the application. In an alternative configuration, the diluent is introduced directly into the pairing disk and allowed to diffuse radially upward, or directly into another disk positioned above the pairing disk.

[0045] The choice of diluent depends on the target of the separation process and the properties of the cell concentrate to be diluted. In some cases, the diluent can be a simple isotonic buffer or deionized water. It may also be advantageous to use a diluent specific to the characteristics of the cell concentrate. For example, in production-scale batch cell cultures with low cell viability, flocculants are typically added to the medium. This is because, when fed to a centrifuge, cells and cell debris aggregate into larger particles, facilitating separation by increasing the sedimentation velocity. Because both cells and cell debris have negative surface charges, compounds used as flocculants are typically cationic polymers, such as polyethyleneimine, that carry multiple positive charges. Because of their multiple positive charges, these flocculants bind to the negatively charged cells and cell debris, forming large aggregates. The use of such flocculants can also result in undesirable increases in the viscosity of the cell concentrate. Therefore, diluents particularly useful in the present invention are deflocculants, which disrupt the binding that increases the viscosity of the cell concentrate. An example of a deflocculating agent is a high salt buffer, such as sodium chloride solution, at a concentration of 0.1 M to 1.0 M. Other deflocculating agents that are effective in reducing the viscosity of cell concentrates are anionic polymers, such as polymers of acrylic acid.

[0046] For cell concentrates where cell viability must be maintained, choose a diluent that is a shear protectant, such as dextran or Pluronic F-68, which, when used in conjunction with an isotonic buffer, increases cell viability upon ejection from the centrifuge.

[0047] The operation of the exemplary centrifuge shown in Figure 4 is described as follows: During a feed cycle, feed suspension enters the rotating bowl apparatus via feed piping 2100. As the feed suspension enters the central cavity 1520 of core 1510 near bottom flange 1200, centrifugal force propels the feed suspension outward along the top surface of bottom flange 1200 and through holes 1530 in core 1510 into separation chamber 1550.

[0048] Centrifugal liquid collects in the separation chamber 1550, a hollow, generally cylindrical space beneath the upper flange 1300 that surrounds the core 1510. The liquid flows upward from the inlet until it encounters the aperture 1540 between the separation chamber 1550 and the pump chamber 1420 at the top of the separation chamber 1550 adjacent the core 1410, and enters the separation chamber through aperture 1530. Particles, being denser than the liquid, move from aperture 1530 toward the outer wall of the separation chamber 1550 by settling (particle concentrate). When the centrifuge 1000 stops rotating, the particle concentrate moves downward under the influence of gravity and enters the nozzle 2110 of the supply line 2100, where it is discharged by the supply / discharge mechanism 2000.

[0049] During rotation, the separated liquid flows through holes 1540 into the separated liquid pump chamber 1420. Within the pump chamber 1420, the rotating separated liquid impacts the stationary pairing disc 1410, converting the liquid's kinetic energy into pressure, which forces the separated liquid upward through the separated liquid discharge channel within the supply / discharge mechanism 2000 and out the separated liquid discharge pipe 2400.

[0050] The addition of radial fins 1630 to the inner surface 1620 of the cap portion 1610 of the rotating centripetal pump 1400 increases the efficiency of this pump 1400. These fins 1630 impart angular momentum of the rotating device to the separation fluid in the pump chamber 1420, which would otherwise slow down due to friction when the rotating separation fluid strikes the stationary pairing disk 1410. The centripetal pump 1400 provides a superior means of evacuation of separation fluid over mechanical seals because of the gas-liquid interface within the pump chamber 1420. Gas within the pump chamber 1420 is protected from contamination by the external environment by the rotating seal 1700. The separation fluid being evacuated between the pairing disks 1410 does not come into contact with air during either the supply or evacuation process, preventing excessive foaming that often occurs when air is introduced into the cell culture medium during the evacuation process.

[0051] In the centrifuge 1000 configuration shown in Figures 4 and 5, the cell concentrate is discharged by periodically interrupting bowl rotation and feed flow, and then pumping out the cell concentrate that collects along the exterior walls of the separation chamber 1550. This process is known as intermittent processing. When the separation chamber 1550 reaches volumetric capacity, the centrifuge rotation is interrupted. The cell concentrate flows down toward the nozzle 2110 of the feed line 2100 and is collected by pumping it out of the feed line 2100. The centrifuge 1000 is externally valved (not shown) to collect the concentrate in a collection vessel (not shown). If the entire bioreactor batch has not been completely processed, the bowl rotation and feed flow are resumed, and the feed / drain cycle continues until the entire batch has been processed.

[0052] As mentioned above, when the cell culture medium is concentrated, or if the cell culture medium contains a significant amount of cell debris, the process slows down because longer residence times are required to capture smaller debris particles, which requires lower feed flow rates, faster filling of the separation chamber 1550, and frequent and repeated interruptions to rotation between culture batches. Additionally, as the cell concentrate becomes more viscous, gravity becomes less effective and the cell concentrate does not reach the bottom of the centrifuge 1000, resulting in longer times and possibly the need for washing to remove any remaining cells.

[0053] Modifications of the exemplary disposable centrifuge configurations shown in Figures 6-13 allow for higher average sedimentation rates without increasing angular velocity, allow the centrifuge 1000 to operate continuously or semi-continuously, and allow diluent to be added to the cell concentrate during cell removal, resulting in easier and more complete cell removal.

[0054] The configuration of the disposable centrifuge structure 1000 shown in Figures 6-12 operates as previously described. Feed suspension is fed into the disposable centrifuge structure 1000 through a feed tube 2100. As the feed suspension impacts the accelerator vanes 1560, these vanes 1560 impart an angular velocity to the feed suspension that approximates the angular velocity of the disposable centrifuge 1000. The use of vanes 1560, rather than holes 1530, allows a larger volume of feed suspension to be delivered to the separation chamber 1550 at a slower radial velocity, thereby avoiding the jetting that can occur when feeding a feed suspension through holes 1530 with openings that have a smaller cross-section than the openings between the vanes 1560. This slow velocity of the feed stream as it enters the separation zone or pool minimizes disturbance of the pool liquid, allowing for more efficient settling.

[0055] As the centrifuge 1000 rotates, particles denser than the separation liquid are forced to the outside of the separation chamber 1550, while the particle-free separation liquid approaches the core 1510. The centrifuge bowl 3100 is shaped like an inverted truncated cone, with a smaller radius at the top than at the bottom. Centrifugal force causes particles to collect in the upper, outer portion of the separation chamber, away from the center. This centrifuge 1000 performs a semi-continuous discharge of the concentrate. When discharging the separation liquid, the operation is generally performed as described with reference to FIG. 4. Discharging the cell concentrate is performed in a similar manner, except that the cell concentrate that collects near the upper outer portion of the separation chamber 1550 flows into the concentrate discharge pump chamber 4400 through a hole 4540 near the upper outer wall of the separation chamber 1550.

[0056] The suspension feed rate and angular velocity of rotation can be monitored using sensors, such as, but not limited to, the vibration sensors disclosed in U.S. Patent No. 9,427,748 (incorporated herein in its entirety). Such a sensor system allows filling at a lower flow rate until the sensor configuration indicates the centrifuge is nearly full, and then the feed rate and angular velocity are adjusted appropriately in response to this information. For example, once the centrifuge is nearly full, the feed rate can be reduced or the feed can be interrupted, and the angular velocity can be increased to increase the settling rate; once settling and draining are essentially complete, the cycle can be repeated. When the system is optimized as described above to minimize the need for process interruptions, it can be operated continuously or nearly continuously at the angular velocity required for settling.

[0057] In the case of semi-continuous concentrate discharge, a concentrate pump 4400 that operates intermittently and removes concentrate is used to continuously pump the suspension into the centrifuge 1000. The operation of the concentrate pump 4400 can be controlled by an optical sensor in the concentrate discharge line that indicates whether concentrate is being discharged. Instead of the concentrate pump 4400, the discharge cycle can be managed using a controller and sensor that determines when to open and close valves to most efficiently process the suspension.

[0058] Furthermore, the average rate of discharge can be controlled by using a centrifuge 1000 with an adjustable gap between the pairing discs 4410, 1410. It is sufficient for the pairing discs 4410, 1410 to be adjustable. Opening the gap between the pairing discs 4410, 1410 (which form part of the flow path exiting the centrifuge 1000) allows flow, while closing it prevents flow. This gap acts as an internal valve. Depending on the desired product or product properties, it can be advantageous to widen or narrow the gap 4415, 1415 between the pairing discs 4410, 1410. Changing the gap affects both the pumping action and the shear rate associated with the pairing discs.

[0059] The rate (volume) of concentrate and centrate removal from the centrifuge 1000, and the viability of the removed concentrate, can be further controlled by utilizing features of the exemplary configurations shown in Figures 4-13. The addition of accelerator fins 4630, similar to those in the centrate pumping chamber 1420, to the concentrate pumping chamber 4420 increases the rate at which concentrate is removed by eliminating some of the frictional slowdown between the flowing concentrate and the disk 4410. In addition to accelerator fins 4630 on the top surface of the pumping chamber 4420, such fins 4630 can also be added to the bottom surface of the pumping chamber 4420 for added effectiveness. Slits can also be used in place of holes 1540, 4540 to minimize shear forces on the material entering the pumping chambers 1420, 4420.

[0060] When concentrate viability is a concern, the placement of a rotatable pairing disk 4410 within the pump chamber 4420 minimizes shear forces on the concentrate while in contact with the surface of the pairing disk 4410. Adjusting the rotational speed of the pairing disk 4410 to a speed between the settling speed and the rotational speed of the separation chamber 1550 balances concentrate viability and discharge rate. The desired angular velocity can be controlled by a number of mechanisms known to those skilled in the art. An example of a control means is a slip clutch, which rotates the pairing disk at an angular velocity that is a fraction of the angular velocity of the centrifuge. The use of a slip clutch is known to those skilled in the art. Additionally, other means for adjusting angular velocity are readily apparent to those skilled in the art.

[0061] A peristaltic pump 2510 can also be used to make the removal of concentrate more efficient and reliable, especially when the feed suspension is highly concentrated. The use of a peristaltic pump 2510 allows for more precise control of the concentrate flow rate from the centrifuge 1000 than using a centripetal pump 4400 alone, as the speed of the centripetal pump is not as easily adjustable as the speed of the peristaltic pump 2510.

[0062] Additionally, a diluent pump 5150 can be used to pump a diluent, such as sterile water or a buffer solution, through the diluent flow path 5000 into the concentrate pump chamber 4420 to reduce the viscosity of the concentrate. See above for a more complete description of useful diluents. The operating speed of either or both of the centripetal pump 2510 and the diluent pump 5150 can be controlled by a concentration sensor 4430 responsive to an automatic controller (described below) located in the concentrate outlet connection 2500. The controller can be programmed to start, stop, or change the pump speed for both diluent addition and concentrate withdrawal in response to the particle concentration of the concentrate and / or in response to standard feed / drain cycles, respectively, in response to the concentration sensor 4430.

[0063] Figure 16 shows another exemplary core configuration for use in a centrifuge that provides continuous feeds of concentrate and centrate for continuous separation. Core 10 is similar to the cores described above that are installed within the rotatable bowl of the centrifuge. During operation, the centrifuge bowl and core rotate about axis 12. The centrifuge includes a stationary device 14 and a rotatable device 16.

[0064] As with the previous embodiment, the settling apparatus 14 includes a supply line 18. The supply line 18 is coaxial with the axis 12 and terminates in an opening 20 adjacent the bottom of the core separation chamber or cavity 22. The settling apparatus further includes a centrifugal pump 24. The centrifugal pump 24, described in detail below, has an inlet opening 26 and an annular outlet opening 28. The annular outlet opening is provided with a separate liquid line 18. The annular outlet opening is fluidly connected to a separate liquid line 30. The separate liquid line extends coaxially around and surrounds the supply line 18.

[0065] In this exemplary configuration, the centrate centripetal pump 24 is located within a centrate pump chamber 32. The centrate pump chamber is part of a rotatable device and is defined by a wall that provides an inlet opening 26 to the centrate centripetal pump that is exposed to a pool of centrate during operation.

[0066] The exemplary configuration further includes a concentrate centripetal pump 34. The concentrate centripetal pump 34 of the exemplary configuration may include a configuration similar to that described in detail above. In the exemplary configuration, the concentrate centripetal pump 34 includes an inlet opening 36 located within a wall that defines an annular periphery of the centripetal pump. Note that the outer diameter of the concentrate centripetal pump 34 is larger than the outer diameter of the centrate pump. The concentrate pump further includes an outlet opening 38. The annular outlet opening 38 is fluidly connected to a concentrate outlet line 40, which extends coaxially around the concentrate line 30.

[0067] In an exemplary configuration, the concentrate centripetal pump inlet opening 36 is located within a concentrate pump chamber 42. The concentrate pump chamber is defined by the rotatable device 16. During operation, the concentrate centripetal pump inlet opening 36 is exposed to the concentrate within the concentrate pump chamber 42. The concentrate pump chamber 42 is vertically bounded by an upper portion 44. At least one fluid seal 46 extends between the outer periphery of the outlet tubing 40 and the upper portion 44. The exemplary seal 46 is configured to minimize the risk of fluid leakage from within the separation chamber and to prevent the introduction of contaminants from areas outside the core.

[0068] During operation of the centrifuge, the bowl and core with the cavity and separation chamber rotate in a rotational direction about axis 12. This rotation in a rotational direction operatively separates the cell suspension introduced via supply line 18 into a centrate, which is discharged via centrate line 30, and a concentrate, which is discharged via retentate outlet line 40.

[0069] A cell suspension enters the separation chamber 22 through a tubing opening 20 at the bottom of the separation chamber. The cell suspension is forced outward by centrifugal force and a plurality of accelerator vanes 48. As the accelerator vanes force the suspension outward, centrifugal force acts on the cell suspension, causing the cell fluid to flow outward toward an annular tapered wall 50 that defines the outer boundary of the separation chamber. As shown, the concentrated cell fluid is forced outward and upward against the tapered wall 50, flowing through a plurality of concentrate slots 52. The concentrate flows upward beyond the concentrate slots and into the concentrate pump chamber 42, from which it is removed by the concentrate centripetal pump 34.

[0070] In the exemplary configuration, in operation, cell-free separation fluid is located adjacent to the vertical annular wall 54 that bounds the interior of the separation chamber 22. This separation fluid flows upward into separation fluid holes 56 in the annular base structure that bounds the separation fluid pump chamber 32, forming a pool of separation fluid within the separation fluid chamber. From the separation fluid chamber, operation of the separation fluid centripetal pump 24 causes the separation fluid to flow and be pumped out of the core via the separation fluid tubing 30.

[0071] In the exemplary configuration of Figure 16, the concentrate and centrate pump configurations are generally the same as in Figure 17. In Figure 17, the centrate centripetal pump 24 is shown in an isometric view. As shown in Figure 17, the exemplary centrate centripetal pump has a disk-shaped body that includes a first plate 58 and a second plate 60. In operation, the first and second plates are held in releasable engagement by fastening members such as screws 62. Of course, in other configurations, other configurations and fastening methods can be used.

[0072] In the exemplary configuration, the second plate 60 includes walls that bound three sides of the curved spiral channel 64. Note that in the exemplary configuration, the centripetal pump includes a pair of generally opposed spiral channels 64. Other configurations may use different numbers and configurations of spiral channels.

[0073] In the exemplary configuration, the first and second plates comprise a disk-shaped body of a centripetal pump having a vertically extending outer wall 67 that defines an annular periphery 66. An inlet opening 68 to the spiral flow passage 64 extends from the annular periphery. An annular collection chamber 70 extends radially outward from the axis 12 and is fluidly connected to the spiral flow passage. The annular collection chamber 70 receives fluid entering through the inlet opening 68. The annular collection chamber 70 is fluidly connected to an annular outlet opening that is coaxial with the axis 12. In this exemplary configuration of the centrifugal pump, the annular outlet opening is an annular space extending between the outer wall of the supply pipe 18 and the inner wall of the second plate 60, with an outlet in the inner wall of the second plate 60 fluidly connected to the centrate outlet pipe 30.

[0074] In the exemplary configuration, each spiral channel 64 is configured to curve in the direction of rotation of the bowl and separation chamber. The direction of rotation is indicated by arrow R in FIG. 17 . In the exemplary configuration, each vertically extending wall 74 that bounds the spiral channel and faces the direction of rotation is curved in the direction of rotation. The curved configuration of the wall 74 that horizontally bounds the spiral channel enhances the pumping characteristics of this exemplary configuration. Furthermore, the opposing wall of each spiral channel in the exemplary configuration also has a similar curved configuration. The curved configuration of the vertically extending wall that horizontally bounds the spiral channel results in a constant cross-sectional area from the inlet of each spiral channel to the collection chamber. This constant cross-sectional area could also be achieved using a generally flat wall 78 that extends between the walls 74, 76 and horizontally bounds the spiral channel on one side. Additionally, in the exemplary configuration, the first plate 58 has a generally planar circular surface 80 on one side that faces inward when the plate is configured into the disk-shaped body of the centripetal pump. In this exemplary configuration, the circular surface 80 provides a horizontal boundary with both sides of the volute channel 64 of the centripetal pump.

[0075] It should be noted that in the exemplary configuration with a pair of plates, one plate may be formed with a recess having walls that bound three of the four sides of the curved spiral channel, and the other plate may be formed with a surface that bounds the remaining side of the spiral channel, and other configurations and structures may be employed in other configurations.

[0076] The exemplary centripetal pump design shown in Figure 16 allows for greater liquid displacement than a paring disk type centripetal pump of the same size. Additionally, this exemplary configuration requires less liquid heating than an equivalent paring disk.

[0077] In this exemplary configuration, the outer diameter of the annular periphery of the centrate centripetal pump 24 is smaller than the outer diameter of the concentrate centripetal pump 34. Using this configuration, the centrate centripetal pump does not remove excessive liquid from the pool of centrate that forms in the centrate pump chamber 32. The presence of sufficient liquid in the centrate pump chamber prevents waves from forming in the centrate adjacent the inlet of the centrate centripetal pump. Waves caused by insufficient liquid can cause vibrations in the centrifuge and core, as well as degrade other performance characteristics.

[0078] The large annular circumference of the concentrate pump in the exemplary configuration allows the concentrate centripetal pump to preferentially pump liquid out of the core. The exemplary configuration allows for control of the ratio of centrate flow to concentrate flow from the core for downstream flow in the concentrate output line 40.

[0079] In exemplary configurations utilizing centripetal pumps with the above configuration, the centrifuge characteristics and flow characteristics can be tailored to the specific feedstocks and requirements of the separation process being performed. Specifically, the circumferential diameter of the centripetal pump can be tailored to optimize the characteristics for a specific process activity. For example, a larger circumferential diameter of the centripetal pump results in stronger flow and higher pressure at the outlet. Furthermore, a larger diameter results in a higher mixing ratio than a smaller diameter. However, a larger diameter results in more heating than a smaller circumferential diameter centripetal pump. Thus, a smaller diameter circumferential diameter can be used to minimize heating. Note that the size, area, and number of outlet openings can be varied, and different spiral flow configurations can be employed, to tailor the flow and pressure characteristics to a particular separation process.

[0080] FIG. 19 is a schematic diagram of an exemplary system useful for maintaining a positive pressure within a separation chamber, referred to herein as a cavity, during cell suspension processing. As discussed in connection with the previous configuration, it is generally desirable to maintain a positive pressure above atmospheric pressure within the separation chamber throughout the process. This configuration reduces the risk of introducing contaminants into the separation chamber by penetrating one or more fluid seals that extend between the stationary device and the rotatable device of the core during processing. Furthermore, as discussed above, it is generally desirable to maintain a positive pressure of air within the separation chamber in contact with the inner surface of the fluid seal. The presence of an air pocket adjacent to the seal prevents the seal from contacting the workpiece, reducing the risk of introducing contaminants into the processed fluid or of fluid leaking from the separation chamber.

[0081] In the exemplary system described with reference to Figure 19, a constant positive pressure is maintained within the separation chamber, thereby reducing the risk of introducing contaminants or leaking other treated liquids.

[0082] As shown generally in Figure 19, the centrifuge includes a rotatable bowl 82. The centrifuge bowl is rotatable about an axis 84 by a motor 86 or other suitable device.

[0083] The exemplary centrifuge configuration shown includes a rotatable disposable core 88 that bounds a cavity 90, referred to herein as the separation chamber.

[0084] As with the other configurations described above, this exemplary core includes a static device comprising a suspension inlet supply line 92 having an inlet opening 94 located adjacent a bottom region of the cavity. The static device further includes at least one centripetal pump 96. The centripetal pump of this exemplary configuration has a disk-shaped body with at least one pump inlet 98 adjacent its periphery and a pump outlet 100 located adjacent the center of the centripetal pump. The pump outlet is fluidly connected to a separator outlet line 102, which extends coaxially around the suspension inlet line in the manner previously described. A rotatable upper portion 104 of the fluid-containing separation chamber is operatively connected to at least one seal 106 that fluidly seals the core cavity relative to the inlet and outlet lines. The at least one seal 106 extends in operative sealing relationship between the annular outer surface of the stationary centrate outlet pipe 102 and a rotatable upper portion 104 of the core having an upper inner wall that internally bounds the cavity 90, as shown.

[0085] In the exemplary configuration, inlet tubing 92 is fluidly connected to pump 108. In the exemplary configuration, pump 108 is a peristaltic pump, which is useful for delivering the cell suspension without damaging it. Of course, this type of pump is merely exemplary, and other types of pumps can be used in other configurations. Furthermore, in the exemplary configuration, pump 108 is reversible, so that pump 108 can operate as a feed pump, delivering a controlled feed rate of the cell suspension through inlet line 110 to the inlet tubing. In a further exemplary configuration, pump 108 can also operate as a concentrate draw / extract pump after the cell concentrate has been separated by centrifugation. In performing this function, pump 108 discharges the cell concentrate from the separation chamber by reversing the fluid flow in inlet tubing 92 from the flow that delivered the cell suspension to the separation chamber. The cell concentrate is then discharged into concentrate line 112. As shown in FIG. 19 , inlet line 110 and concentrate line 112 are selectively opened and closed by valves 114 and 116, respectively. In an exemplary configuration, these valves 114 and 116 comprise pinch valves to open and close flow in flexible lines or tubing, although of course this method is exemplary only and other methods may be used in other configurations.

[0086] In an exemplary system, the centrate outlet tubing 102 is fluidly connected to a centrate discharge line 118. The centrate discharge line is fluidly connected to a centrate discharge pump 120. In an exemplary configuration, the centrate discharge pump 120 is a variable flow rate pump that selectively adjusts the flow rate. For example, in some exemplary configurations, the pump 120 may comprise a peristaltic pump having a motor whose speed can be controlled to selectively increase or decrease the flow rate of the pump. The outlet of the centrate discharge pump delivers treated centrate to a suitable collection chamber or other processing device.

[0087] 19, the pressure damping reservoir 122 is fluidly connected to the centrate outlet line 118, which is fluidly intermediate the centrate outlet piping 102 and the pump 120. In the exemplary configuration, the pressure damping reservoir comprises a generally vertically extending vessel that holds the centrate in a fluid-tight relationship within its interior region. The pressure damping reservoir includes a bottom port 124 that is fluidly connected to the centrate outlet line 118.

[0088] Opposite the reservoir 122 is a top port 126. This top port is exposed to air pressure. In an exemplary configuration, the top port is exposed to air pressure from a high-pressure air source, generally designated 128. In an exemplary configuration, the high-pressure source comprises a compressor, air storage tank, or other suitable device that provides a source of high-pressure air above atmospheric pressure within the range required for system operation. Air from the high-pressure air source 128 is filtered through a sterilizing filter 130 to remove impurities. A regulator 132 maintains a generally constant above-atmospheric air pressure level at the top port of the pressure damping reservoir. In an exemplary configuration, the air pressure regulator comprises an electronic, fast-acting regulator, thereby ensuring that a generally constant air pressure is maintained at the desired level. The exemplary fast-acting regulator 132 activates immediately, increasing the pressure acting on the top port 126 when the pressure drops below the desired level, and immediately releases the regulator pressure when the pressure acting on the top port rises above the regulator setting.

[0089] In some configurations, the regulator outlet may be operatively fluidly connected to the interior of the upper portion 104 of the separation chamber via an air line 143, shown generally in phantom. In this exemplary configuration, regulator outlet pressure acting on the reservoir upper port 126 also acts via the air line 143 on an air pocket within the separation chamber. This air pocket extends downward to the level of the cavity above the peristaltic pump inlet, into the interior of the at least one seal 106, and radially from an area adjacent the axis 84 to the interior upper wall of the upper portion 104. In the exemplary configuration, the line 143 applies positive pressure to an area within the separation chamber below the at least one seal via at least one separate flow path. This separate flow path extends through the stationary structure of the apparatus, including the centrate outlet piping 102 and the inlet supply piping 92. The at least one separate flow path of the air line 143 applies air pressure to the interior of the upper portion 104 via at least one air opening that opens into the separation chamber. The at least one opening 145 is located on the exterior of the outlet pipe 102 above the inlet 98 to the centripetal pump and below the at least one seal 106. Of course, this configuration of the air lines applying positive air pressure to the air pocket of the separation chamber and inside the at least one seal is exemplary only, and other configurations and methods can be used in other configurations.

[0090] In the exemplary configuration of pressure damping reservoir 122, upper level sensor 134 detects the separated liquid within the pressure damping reservoir. The upper level sensor operationally detects the separated liquid at the upper level. The lower level sensor 136 detects the liquid in the reservoir at a lower level. The upper level sensor 138 is positioned to detect a high level in the reservoir above the upper level. This high level sensor detects an unacceptably high level, indicating an abnormal condition requiring system shutdown or other safety measures. In the exemplary configuration, level sensors 134, 136, and 138 comprise capacitive proximity sensors useful for detecting adjacent separated liquid levels within the pressure damping reservoir. These types of sensors are exemplary only, and other sensors and methods may be used in other configurations.

[0091] The exemplary configuration may further include other components appropriate to the operation of the system. For example, other valves, lines, pressure connections, and other suitable components may be included to accommodate the processing and operation of the suspension, the centrate, the concentrate, and the like, as appropriate for the particular system. For example, an additional valve may be valve 140 for controlling the open / closed state of the centrate discharge line 118. The additional lines, valves, connections, and components that may be used in the system will vary depending on the nature of the system.

[0092] The exemplary system of FIG. 19 further includes at least one control circuit 142, sometimes referred to as a controller. The at least one control circuit 142 includes one or more processors 144. The processor is operatively connected to one or more data storage devices 146. Here, a processor refers to an electronic device that processes data stored in one or more data storage devices or received from external sources, analyzes information, controls other devices, or performs other actions in accordance with processor-executed instructions. The one or more control circuits may be implemented as hardware circuits, software, firmware, or applications, enabling the control circuit to receive, store, or process data and perform other actions. For example, the control circuit may be implemented as one or more microprocessors, CPUs, FPGAs, ASICs, or other integrated circuits or other types of circuitry capable of performing functions in the manner of an electronic computing device. Note that the data storage device may correspond to one or more volatile or non-volatile memory, such as RAM, flash memory, hard drives, solid-state devices, CDs, DVDs, optical memory, magnetic memory, or other circuit-readable media for storing computer-executable instructions and / or data.

[0093] The circuit-executable instructions may include any of a number of programming languages ​​and formats, including, but not limited to, routines, subroutines, threads of execution, objects, scripts, methodologies, and functions that perform actions such as those described herein. The control circuitry architecture encompasses, corresponds to, and utilizes principles described in the textbook entitled "Microprocessor Architecture, Programming, and Applications with the 8085" by Ramesh S. Gaonker (Prentice Hall, 2002), which is incorporated herein by reference. Of course, these circuit architectures are exemplary only, and other configurations may use other circuit architectures that store, process, analyze, and output information.

[0094] In exemplary configurations, the at least one control circuit 142 is operatively connected to a suitable interface with at least one sensor, such as sensors 134, 136, and 138. The at least one control circuit is also operatively connected to variable flow displacement pump 120. Additionally, in some exemplary configurations, the at least one control circuit may be operatively connected to other devices, such as motor 86, pump 108, regulator 132, air pressure source 128, fluid control valves, and other devices.

[0095] At least one of the exemplary control circuits described above operatively receives data and controls such devices in accordance with circuit-executable instructions stored in data storage device 146. In an exemplary configuration, the fluid level 147 in the fluid damping reservoir is characteristic of the pressure in the centrate discharge line 102. One exemplary configuration that does not use air line 143 utilizes the fact that the pressure in the centrate discharge line indicates the pressure in the upper core section 104 and, in turn, the pressure in the separation chamber adjacent seal 106 to control the operation of the discharge pump and other components. As previously discussed, it is desirable to maintain a positive pressure above atmospheric pressure and an air pocket adjacent at least one seal in the separation chamber to prevent the introduction of impurities into the separation chamber resulting from negative pressure. Note that if the fluid level in the separation chamber becomes too high, the pressure and the suspension being treated will overflow the seal, creating potential contamination issues and undesirable exposure and loss of the treated liquid. This is due to excessive backpressure on the centrate line connecting to the outlet from the centripetal pump.

[0096] In the exemplary configuration, the rotational speed of the bowl determines the pumping force and pump output pressure level of the centripetal pump. This pressure output level of the centripetal pump varies with the rotational speed of the bowl and core. In the exemplary configuration without air line 143, back pressure should be controlled in the centrate outlet piping. Back pressure is generated by controlling the speed of the motor operating pump 120 and pressure damping reservoir level 147. Back pressure is maintained below the pump output pressure (allowing the centripetal pump to pump centrate from the separation chamber) and a positive pressure above atmospheric pressure to ensure that contaminants do not pass the seal into the separation chamber, maintaining high pressure air in the separation chamber adjacent to the seal and isolating the seal from the components of the suspension being processed.

[0097] In an exemplary configuration, the high pressure applied to the upper port 126 of the pressure damping reservoir is maintained by a regulator 132. Additionally, at least one control circuit 142, which maintains the speed of the pump 120 and maintains a liquid level 147 between an upper liquid level 134 and a lower liquid level 136 detected by a sensor 134, controls the flow of separated liquid from the separation chamber, maintaining a desired constant pressure in the upper region of the separation chamber and preventing separated liquid from contacting or overflowing the seal.

[0098] In an alternative configuration using air line 143, the positive pressure level from the regulator acts on both the fluid in reservoir 122 and the area of ​​the separation chamber above the centripetal pump inlet. Because the positive pressure level is the same at both locations, the back pressure in the centrate discharge line (the pressure exerted on the fluid in the reservoir) is nearly always the same as the pressure in the air pocket above the separation chamber. This allows the centripetal pump to operate completely unaffected by either pressure.

[0099] In this exemplary configuration, pump 120 and other system components are controlled in response to at least one control circuit 142. This ensures that a sufficient volume of air is present within reservoir 122 at all times during centrate production. The reservoir therefore provides a desired damping effect to dampen pressure changes in the centrate discharge line that would otherwise result from the pumping action of pump 120. Damping is achieved by maintaining the liquid in reservoir 122 below an upper liquid level detected by sensor 134. Furthermore, the reservoir liquid level is controlled to remain above a lower liquid level detected by sensor 136. This ensures that the centripetal pump does not expel air, thereby minimizing aeration in the centrate.

[0100] In an exemplary configuration, the flow of clarified liquid from the separation chamber is controlled by operation of at least one control circuit. The exemplary control circuit operates the system under processing conditions to maintain a constant overall flow rate of the cell suspension into the separation chamber 90 using pump 108, while operating motor 86 to perform the separation process, maintain a constant bowl speed, and separate the clarified liquid from the cell concentrate. The exemplary configuration also operates to maintain an ideal constant back pressure on the clarified liquid discharge line from the centripetal pump, while maintaining air in the separation chamber above the level of the lower air pocket, separating at least one seal 106 from the clarified liquid and the concentrate being processed.

[0101] In an exemplary configuration, the pressure maintained through manipulation of the regulator in the pressure damping reservoir is set to approximately 2 kPa (0.29 psi) above atmospheric pressure. In the exemplary system, this pressure setting has been found to ensure that seal integrity and isolation are maintained throughout all stages of cell suspension processing. Of course, this setting is exemplary only, and other configurations may employ other pressure settings, pressure damping reservoir configurations, sensors, etc.

[0102] FIG. 20 is a schematic diagram illustrating exemplary logic implemented through operation of at least one control circuit 142 in maintaining a desired pressure level in the centrate discharge line and in the upper portion of the separation chamber. It should be noted that the control circuit in some exemplary configurations may perform numerous additional or different functions beyond those described above, including the pressure control functions described above as well as overall control of different processes and steps in the operation of the centrifuge. As shown in FIG. 20, an initial subroutine, step 148, may operate at least one control circuit 142 to determine whether the centrifuge is currently in a mode of operation in which centrate is being discharged from the separation chamber. If so, the at least one control circuit operates to operate the centrate discharge pump 120 to discharge centrate from the centrate discharge line 118. This is accomplished by operating the pump motor. In exemplary configurations, the flow rate of the pump 120 may be initially set to a set value or may be varied depending on specific operating conditions, which may be determined through operation of the control circuit during the process. Operation of the centrate discharge pump is performed in step 150.

[0103] Next, at least one control circuit operates to determine whether liquid is at a high level as indicated by high liquid level sensor 138 in step 152. This level indicates an undesirable condition. If liquid is detected at sensor 138, the control circuit operates to take steps to address the condition. This may involve operating pump 120 to increase the flow rate, continuing centrifuge operation, and determining whether the liquid level drops within a predetermined time. Alternatively, or in addition, at least one control circuit may slow down pump 108 to reduce the flow of incoming liquid. If these actions do not result in a drop in the liquid level within the predetermined time, additional steps may be taken. These steps may include slowing or stopping the rotation of bowl 182. These actions may include stopping operation of pump 108 to avoid introducing more suspension into the separation chamber. These steps, commonly referred to as shutting down normal operation of the system, may be identified as step 154.

[0104] If no liquid is detected at the upper level sensor 138, the at least one control circuit operates to determine whether liquid is detected at the upper level sensor 134. This is step 156. If liquid is detected at the upper level, the at least one control circuit operates in response to its stored instructions to increase the speed, and therefore the flow rate, of the discharge pump 120. In the exemplary configuration, this is done by increasing the speed of a motor that is part of the pump. This is step 158. As the pump flow rate is increased, the pressure damping reservoir liquid level 147 begins to drop as more liquid is displaced by the pump 120.

[0105] If liquid is not detected at the upper level of sensor 134 in step 156, at least one control circuit operates to determine whether liquid is detected at the upper level of sensor 134. This is step 160. If liquid is not detected at the upper level, at least one control circuit operates according to its program to reduce the flow rate of pump 120. In exemplary configurations, this is done by reducing the motor speed. This is step 162. In exemplary configurations, reducing the flow rate of pump 120 begins to increase the liquid level 147 in the pressure damping reservoir. In some exemplary configurations, if the liquid level does not increase in the reservoir within a predetermined time, the control circuit operates according to its program to take additional measures, such as the stop step 154 ​​described above. The control circuit in exemplary configurations operates to vary the supply rate of pump 120 to maintain the liquid level 147 in the pressure damping reservoir at a generally constant level between the liquid levels of sensors 134 and 136 during separation liquid production.

[0106] The exemplary configuration maintains a generally constant high pressure of sterile air throughout the liquid in the pressure damping reservoir, thereby ensuring consistent maintenance of a similar high pressure in the centrifugal outlet piping and at the seals within the separation chamber. The exemplary configuration also maintains a desired pressure level during different operating conditions of the centrifuge, including conditions during which the bowl rotates at different speeds. These conditions include, for example, conditions during the initial filling of the separation chamber with a cell suspension at a relatively high flow rate and conditions during which the centrifuge rotates at a relatively slow speed. Pressure can also be maintained during the subsequent final filling conditions, during which the cell suspension flow rate into the separation chamber is lower and the bowl rotation speed is higher. Furthermore, positive pressure can be maintained as described above during the supply of suspension to the bowl and the discharge of centrifugal liquid from the separation chamber. The exemplary configuration also operates at least one control circuit to maintain positive pressure during the discharge of concentrate from the separation chamber. Maintaining positive pressure within the separation chamber throughout these conditions reduces the risk of contamination and undesirable conditions that would otherwise result from negative (subatmospheric) pressure conditions.

[0107] Of course, the above features, components, structures, and control methods are merely exemplary, and other approaches may be used in other configurations. Furthermore, although the system in the exemplary configuration processes both the permeate and concentrate in a batch mode rather than a continuous mode, the principles are applicable to other types of systems.

[0108] While the use of a pressure damping reservoir in the exemplary configuration ensures that desired pressure levels are maintained in the outlet tubing and separation chamber, other exemplary configurations may use other methods. For example, in some configurations, pressure may be directly sensed and / or applied to the outlet tubing, separation chamber, or other location corresponding to the pressure in the separation chamber. In some configurations, the flow rate of the discharge pump may be controlled to maintain a desired pressure level. In yet other configurations, the exemplary control circuitry may operate to control both the discharge pump and a pump that supplies the suspension to the core and / or suitable valves or other flow control devices to maintain a desired pressure level. The selection of such alternative approaches may be dependent on the particular centrifuge being used and the liquid being processed, among other factors.

[0109] 21 is a schematic diagram illustrating another centrifuge system 170 configured for continuous or semi-continuous separation of a cell culture batch into a cell detachment and a cell concentrate. The exemplary system includes a rigid centrifuge bowl 172 that is rotatable about an axis 174. The bowl includes a cavity 176 configured to releasably receive a disposable structure 178. The rigid bowl includes an upper opening 180. An annular retaining ring or other retaining structure 182 releasably secures the disposable structure 178 within the bowl cavity.

[0110] The exemplary disposable structure 178 of this configuration includes a supply line 184 extending along a central axis. As described below, the supply line is used to deliver cell culture batch fluid to an interior region 186 of the disposable structure 178. The supply line 184 extends from an upper portion at a first axial end 188 of the disposable device to a lower portion at a second axial end 192 of the disposable structure 178 to the interior region. The disposable structure 178 includes a generally disk-shaped portion 194 adjacent the first axial end. The exemplary disk-shaped portion 194 is generally rigid, i.e., rigid or semi-rigid, and includes an annular periphery 196 that is configured to engage the annular boundary wall 198 of the centrifuge bowl cavity 176. The annular periphery of the disk-shaped portion 194 is configured to engage the rigid bowl 172 so that the disposable structure rotates therewith.

[0111] Additionally, the exemplary disposable structure 178 includes a hollow, rigid or semi-rigid, cylindrical core 200 that operatively engages and rotates unitarily with the disk-shaped portion 194. The core 200 is axially aligned with the disk-shaped portion and extends axially midway between the top and bottom of the disposable structure 178. The core 200 includes an upper opening 202 and a lower opening 204 through which the supply tubing 184 passes.

[0112] The disk-shaped portion 194 includes a substantially circular centrifugal pump chamber 206. A centrifugal pump 208 is located within the pump chamber 206. A substantially annular centrifugal opening 210 is fluidly connected to the centrifugal pump chamber 206. "Substantially annular" here means that the opening may be discrete and / or continuous in an annular configuration. The centrifugal pump 208 is fluidly connected to a centrifugal discharge line 212. The centrifugal discharge line 212 extends coaxially and surrounds the supply line 184. Discharged centrate passes through the substantially annular opening on the periphery of the centrifugal pump and the annular space of the centrifugal pump discharge line 212, which is external to the supply line.

[0113] Additionally, disk-shaped portion 194 includes a concentrate centripetal pump chamber 214. The concentrate centripetal pump 214 is a generally cylindrical chamber located above the centrate centripetal pump chamber 206. The concentrate centripetal pump chamber 214 has a concentrate centripetal pump 216 located therein. The concentrate centripetal pump is fluidly connected to a concentrate discharge line 220. The concentrate discharge line 220 extends in an annular surrounding relationship with the centrate discharge line 212. The concentrate passes through a generally annular opening in the periphery of the concentrate centripetal pump and through an annular space within the concentrate discharge line 220 that is outside the centrate discharge line.

[0114] The generally annular concentrate opening 218 is fluidly connected to the concentrate pumping chamber 214. In an illustrative example, the generally annular concentrate opening and the generally annular separate opening are concentric openings, with the concentrate opening radially outward from the separate opening. Of course, this configuration is illustrative only, and other approaches and configurations are possible in other configurations.

[0115] Additionally, the exemplary disposable structure 178 includes a flexible outer wall 222. The flexible outer wall 222 is a fluid-tight wall that, in the operative position of the exemplary disposable structure 178, extends in supporting engagement with the wall bounding the rigid bowl cavity 176. In the exemplary configuration, the flexible outer wall 222 operatively engages and forms a rigid fluid connection with the disk-shaped portion 194. The flexible outer wall is internally frusto-conical, with a reduced inner radius adjacent the bottom of the disposable structure adjacent the second axial end 192.

[0116] The exemplary flexible outer wall 222 extends in surrounding relationship around at least a portion of the core 200. Further, the outer wall 222 bounds an annular separation chamber 224. The separation chamber 224 extends radially between the outer wall of the core 200 and the flexible outer wall 222. The generally annular concentrate opening 218 and the generally annular separate liquid opening 210 are each in fluid communication with the separation chamber 224.

[0117] In an exemplary configuration, flexible outer wall 222 has a textured outer surface 226. This textured outer surface allows air to escape from the space between flexible outer wall 222 and the surface that bounds the cavity of rigid bowl 172. In an exemplary configuration, the textured outer surface covers substantially the entire area of ​​the flexible outer wall that contacts the rigid bowl. In an exemplary configuration, the textured outer surface has one or more patterns of outwardly extending protrusions or depressions 228 that form spaces or recesses therebetween to facilitate ventilation. Ventilation is achieved in the bowl cavity when disposable structure 178 is placed in the bowl cavity through either top opening 180 or bottom opening 230. In an exemplary configuration, the protrusions may be constructed of a resiliently deformable material that reduces in height in response to the force of the liner against the rigid wall of the bowl. The textured outer surface 226 of the flexible outer wall 222 reduces the likelihood of air pockets being trapped between the rigid bowl of the centrifuge and the disposable structure. Such air pockets can cause variations in the wall contour, potentially imbalances and / or distort the contour of the separation chamber in a manner that adversely affects the separation process. Of course, the air release structures described above are exemplary only, and other air release structures can be employed in other configurations.

[0118] Additionally, the exemplary disposable structure shown in FIG. 21 includes a lower disk-shaped portion 232 that is rigid or semi-rigid. This rigid or semi-rigid material allows it to maintain its shape during operation. In the exemplary configuration, the lower disk-shaped portion 232 is conical and operatively connected to and attached to the lower end of the core portion 200 by a vertically extending wall or other structure. A plurality of angularly spaced channels 234 extend between the upper surface of the disk-shaped portion 232 and the radially outward lower portion of the core portion. Because the channels 232 extend radially outward and upward relative to the bottom of the second axial end 192, cells in the cell culture batch solution entering the interior region 186 through the opening 190 of the supply line 184 flow radially outward and upward into the separation chamber 224.

[0119] In the exemplary configuration, the flexible outer wall 222 extends below the lower disk-shaped portion 232 at the second axial end 192 of the disposable structure. The flexible outer wall 222 extends halfway between the lower disk-shaped portion 232 and the wall of the rigid bowl 172 that bounds the cavity in which the disposable structure is located.

[0120] In the exemplary configuration, the centrifuge bowl 172, upper disk-shaped portion 194, lower disk-shaped portion 232, and flexible outer wall 222 rotate while the supply line 184, the centrate discharge line 212, the concentrate discharge line 220, the centrate centripetal pump 208, and the concentrate centripetal pump 216 remain stationary. At least one annular resilient seal 236 extends between the outer surface of the concentrate discharge line 220 and the upper disk-shaped portion 194 for operative sealing engagement. Because the at least one seal 236 maintains an airtight seal as described above, an air pocket can be maintained within the interior region 186 during cell processing, isolating the seal from the processed cell culture batch. The air pocket maintained within the interior region of the disposable structure maintains the centrate centripetal pump 208 and the concentrate centripetal pump 216 in fluid communication with the cell culture batch. As above, a positive pressure can be maintained within the interior region, ensuring isolation of the at least one seal 236 from the processed cell culture batch. Alternatively, other approaches may be employed to maintain separation of the seal from the liquid being treated.

[0121] Exemplary system 170 operates similarly to that described above. Cells in a cell culture batch enter interior region 186 of disposable structure 178 via supply line 184. The cells enter interior region 186 via supply line opening 190 at the lower axial end of the disposable structure. Centrifugal force causes the cells to flow outward through opening 234 and into separation chamber 224. Due to the outward and upward tapering of outer wall 222, cell concentrate containing cells or cell fluid collects in the radially outward upper region of separation chamber 224. Totally cell-free separation fluid collects in the separation chamber adjacent radially inward to the outer wall of core 200.

[0122] In the exemplary configuration, cell separation fluid flows upward through a generally annular separation fluid opening into the separation fluid pump chamber. The separation fluid passes inward through a generally annular opening in the separation fluid centripetal pump and then flows upward through separation fluid outlet piping 212. Simultaneously, cell concentrate flows through a generally annular outlet opening 218 into the concentrate centripetal pump chamber 214. The cell concentrate passes inward through a generally annular opening in the concentrate centripetal pump 216 and then flows upward through separation fluid outlet piping 220. This exemplary configuration allows for continuous or semi-continuous operation of the exemplary system 170. The ability to control the operation of the system 170 in a similar manner as described above allows the system to operate reliably over extended periods of time and deliver the desired cell concentrate and the generally cell-free separation fluid as separate output fluid streams.

[0123] 22 illustrates another centrifuge system 238. The system 238 includes a disposable structure 240, which is similar in many respects to the previously described disposable structure 178. Some of the structures and features of the disposable structure 240 that are generally the same as the disposable structure 178 described above are designated with the same reference numerals as the disposable structure 178.

[0124] The disposable structure 240 differs from the disposable structure 178 in that it includes a rigid or semi-rigid lower disk-shaped portion 242. The lower disk-shaped portion 242 is a generally conical structure that operatively connects to the lower end of the core portion 200. A plurality of radially outwardly and upwardly extending channels 244 extend between the lower end of the core portion 200 and the lower disk-shaped portion 242. The exemplary lower disk-shaped portion 242 further includes angularly spaced radially extending vanes 246. The channels extend radially outward between each pair of angularly adjacent vanes 246. In this exemplary configuration, the vanes 246 extend upwardly from the bottom of the disk-shaped portion 242, with at least some of the vanes 246 operatively engaging the core portion at their radially outer portions. In this exemplary configuration, the vanes 246 facilitate movement and separation within the interior region of the disposable structure to accelerate cell culture batch processing.

[0125] 23 illustrates another exemplary configuration of a centrifuge system 248. This exemplary configuration includes a disposable structure 250. This disposable structure 250 is similar in many respects to the disposable structure 178 described above. Some of the structures and features of the disposable structure 250 that are the same as those of the disposable structure 178 described above are designated by the same reference numerals.

[0126] Disposable structure 250 differs from disposable structure 178 in that it includes a lower disk-shaped portion 252. Lower disk-shaped portion 252 is a generally rigid or semi-rigid conical structure that is operatively connected to core 200 via a wall or other suitable structure. Lower disk-shaped portion 252 includes a plurality of angularly spaced, radially outwardly extending accelerator vanes 254 that extend downwardly from the lower conical side of disk-shaped portion 252. Each directly and angularly adjacent pair of vanes 254 includes a flow path extending therebetween. In this exemplary configuration, a flexible outer wall 222 extends intermediate the lower ends of vanes 254 and the wall of rigid bowl 172 that bounds cavity 176. In this exemplary configuration, the accelerator is submerged and accelerates the cell culture batch liquid, facilitating separation within the interior region of the disposable structure. Of course, the features of the disposable structures described herein may be combined in different configurations to facilitate the separation and processing of different types of materials and materials with different properties, and to generate a desired output fluid flow.

[0127] Figure 26 illustrates another disposable structure 304. This disposable structure 304 is similar to the disposable structure 178 described above, except as described below. Elements that are the same as those in the disposable structure 178 are identified in Figure 26 using the same reference numerals.

[0128] The disposable structure 304 includes a continuous, annular concentrate dam 306. The concentrate dam 306 extends downwardly into the separation chamber 224 and extends radially inward from the generally annular concentrate opening 218. The exemplary annular concentrate dam, shown in cross section, extends downwardly below the concentrate opening and includes a tapered outer surface 308 that extends outward in the axial cross section toward the opening 218.

[0129] Additionally, the disposable structure 304 includes a continuous, annular separate liquid dam 310 that extends downward into the separation chamber 224 below the generally annular separate liquid opening 210. The separate liquid dam 310 is positioned radially outward from the separate liquid opening 210. In the exemplary configuration, the concentrate dam 306 and the separate liquid dam 310 extend approximately the same downward distance into the separation chamber 224. However, other configurations may be employed. Also, in other configurations, the centrifuge structure may include either a concentrate dam or a separate liquid dam, but not both.

[0130] An annular recess 312 extends radially into the separation chamber between the separate and concentrate dams. This exemplary annular recess extends upward between the separate and concentrate dams, forming an annular pocket therebetween.

[0131] In the exemplary configuration, the concentrate dam 306 ensures that primarily cells and other solids to be separated pass outward along the top surface bounding the separation chamber 224 to reach the concentrate opening 218 and concentrate centripetal pumping chamber 214. Additionally, the centrate dam 310 allows primarily cell-free centrate to flow along the top surface bounding the separation chamber 224 into the generally annular centrate opening 210 and into the centrate pumping chamber 206. Note that numerous variations of the concentrate dam and centrate dam are possible in different configurations depending on the nature of the liquid being treated and the operating conditions for such liquid.

[0132] 24 is a schematic diagram illustrating an exemplary control system for continuously processing a generally cell culture to produce a generally cell-free centrifuge solution and a cell concentrate stream. This exemplary configuration uses the centrifuge system 170 described above. It should be noted that the features of the exemplary system can be used with many different types of feedstocks, centrifuge systems, and configurations described herein.

[0133] In the exemplary configuration shown, centrifuge bowl 172 is rotated at a predetermined speed about axis 174 by motor 256. Supply tubing 184 is operatively connected to a cell culture supply line 258 that receives cell culture batch fluid. This supply line is operatively connected to a supply pump 260. In the exemplary configuration, supply pump 260 may be a peristaltic pump or other pump suitable for delivering cell culture fluid to a disposable structure at a predetermined flow rate.

[0134] Centrate outlet tubing 212 is fluidly connected to a centrate outlet line 262. A centrate optical density sensor 264 is operatively connected to an interior region of the centrate outlet line 262. In an exemplary configuration, the centrate optical density sensor is an optical sensor that operatively determines the density of cells currently exiting the disposable structure. In an exemplary configuration, this can be achieved by measuring a decrease in the intensity of light output from an emitter received by a receiving device and carrying at least a portion of the centrate flow. The amount of light received by the receiving device from the emitter decreases as the cell density of the centrate increases. This is only one example of a sensor that can be used to determine the density or amount of cells present in the centrate; in other configurations, other types of sensors can be employed. For example, the light can be near-infrared light, other visible light, or invisible light. In other detection configurations, electromagnetic, acoustic, or other signal forms can be used. The centrate outlet line is operatively connected to a centrate pump 266. In an exemplary configuration, the centrate pump may be a peristaltic pump or other variable flow pump suitable for pumping centrate.

[0135] In the illustrated configuration, concentrate discharge line 220 is operatively connected to an interior region of concentrate discharge line 268. A concentrate optical density sensor 270 is operatively connected to at least a portion of the interior region of concentrate discharge line 268. The illustrated concentrate optical density sensor may operate similarly to the centrate optical density sensor described above. It should be understood that concentrate optical density sensors may have different structures or characteristics, and that different types of cell density sensors may be utilized in other exemplary configurations. Concentrate discharge line 268 is operatively connected to concentrate pump 272. In the illustrated configuration, concentrate pump 272 may comprise a peristaltic pump or other variable flow pump suitable for pumping concentrate without damaging it. It should be noted that these structures and components are exemplary, and other systems may include different or additional components.

[0136] The exemplary control system includes control circuitry 274, sometimes referred to herein as a controller. In an exemplary configuration, the control circuitry may include one or more processors 276 and may also include one or more data storage devices 278. The one or more data storage devices may include one or more tangible media that store circuit-executable instructions and data that, when executed by the controller, perform the operations described below. Examples of such media include solid-state memory, magnetic memory, optical memory, and other non-transitory media for storing circuit-executable instructions and / or data. Additionally, the control circuitry may include structures such as those described above.

[0137] The operations performed by an exemplary controller 274 will now be described with reference to the logic flow schematic diagram shown in Figure 25. In the exemplary configuration, the controller 274 operates to control the operation of each component in the system to maintain an output stream that simultaneously produces a cell-free centrifuge and a cell concentrate, collectively, by using optical density sensors in the centrifuge and concentrate outlet lines, respectively, to detect the cell density (or turbidity) of the output streams and adjust the operation of the system components to maintain the output within a desired range.

[0138] When using the exemplary control system, the cell concentration in the cell culture fluid to be processed is measured prior to initiating system operation. The desired axial rotation speed of the centrifuge is determined, as is the operating speed of the feed pump 260. In the exemplary configuration, the rotation speed of the centrifuge and the rate at which the feed cell fluid is delivered by the feed pump are generally maintained at constant set points by the controller. Of course, other configurations and systems may employ other methods by which the controller can adjust speeds and flow rates during cell processing.

[0139] In an exemplary configuration, the discharge rate (flow rate) of the external concentrate pump 272 is set to an initial value (sometimes referred to herein as a "prime value") based on the determined cell concentration. In an exemplary configuration, a "prime duration" corresponding to the time the external concentrate pump 272 initially operates at the initial value is also set. During this duration, the disposable structure 178 is partially filled. In this exemplary system, a "basal rate" is set for the concentrate pump based on the cell density as well as the feed rate from the feed pump 260. The basal rate of the concentrate pump is the speed (corresponding to the flow rate) at which the concentrate pump will operate after the prime duration. In an exemplary configuration, the set basal time is considered to correspond to a concentrate pump speed that will produce a centrate with a cell density below a desired set limit and a cell concentrate with a cell density above a desired set range. These set values ​​and ranges are received in response to input from a suitable input device and stored in at least one data storage device.

[0140] 25, operation of the concentrate pump 272 at the initial speed is indicated by step 280. The controller determines whether the concentrate pump has been operating at the initial speed for a time corresponding to an initial duration to at least partially fill the disposable structure 178.

[0141] Once the concentrate pump has run at the initial speed for the initial required time, the controller increases the concentrate pump speed to the base speed as indicated by step 284. The controller 274 operates to monitor the cell density of the permeate as detected by sensor 264. The controller also determines whether the optical density is greater than the desired set point as indicated by step 286. If the optical density of the permeate is not greater than the set point, there is sufficient cells or cellular material in the permeate, so the controller determines no change in the operating speed of the concentrate pump and the logic returns to step 284.

[0142] If, at step 286, the optical density of the permeate is found to be higher than the set point, the logic proceeds to step 288. At step 288, the controller operates to increase the speed of the concentrate pump by a set incremental step amount. The purpose of this speed increment is to cause the overall optical density of the permeate to clear the set point as a result of the reduction in cell count in the permeate.

[0143] After increasing the concentrate pump 272 speed in step 288, the controller, responsive to the sensor 264, determines in step 290 whether the optical density of the centrate is still higher than the set point after a set time has elapsed since the concentrate pump speed (flow rate) increment. If so, the controller continues to monitor the optical density of the centrate until it is no longer higher than the set point. In an exemplary configuration, the instructions include a set time. During this set time, the centrate optical density must not be higher than the set point before the concentrate pump speed controller determines that an adjustment to the base rate is sufficient to maintain the centrate optical density at or below the desired set point. In step 292, the controller determines that the increased concentrate pump speed has maintained the centrate optical density below the set point for a retention set time, which corresponds to either consistently producing a centrate with sufficient cell removal or reaching a programmed wait time. In response to a consistent production of sufficiently cell-cleaned permeate, or in response to reaching a programmed wait time, the controller adjusts the concentrate pump basal rate value to correspond to the increased basal rate in step 294. The controller sets the new basal rate, and the logic returns to step 284. Note that if step 286 determines that the permeate optical density is still above the set point, the concentrate pump speed may be readjusted.

[0144] The example controller simultaneously monitors the optical density of the cells in the output concentrate stream. This can be done by monitoring the optical density detected by sensor 270. As shown in step 296, the controller operates to determine whether the optical density in the concentrate is below a desired set point. If this optical density is equal to or greater than the desired set point value stored in the data storage device, the cell concentration in the concentrate output stream is above the desired level, and the logic returns to step 284. If the optical density of the concentrate is below the desired set point, this means that the cell level in the concentrate is below the desired level, and the controller proceeds to step 298. In step 298, the speed of the concentrate pump is slowed down by a predetermined incremental step. Slowing the concentrate pump reduces the output flow rate, which, overall, reduces the amount of cells in the concentrate output stream, resulting in a higher optical density in the concentrate output stream.

[0145] The concentrate pump 272 then operates at the new reduced speed, as shown in step 300. The controller operates the concentrate pump 272 at this reduced speed for a set time corresponding to a set value stored in the data storage device, as shown in step 302, allowing for a high concentration of cells in the output concentrate stream before making a determination as to whether the reduction in speed is sufficient. After this time has elapsed in step 302, the controller returns to step 284, from which point the logic flow repeats to determine whether further speed adjustments are required.

[0146] Of course, this simplified schematic logic flow is illustrative only, and in other configurations, different logic flows and / or additional operational parameters of system components may be monitored and adjusted to achieve desired output flows of centrate and concentrate. For example, in another illustrative example, the speed of the centrate discharge pump, and therefore the centrate discharge flow, may be varied at least in part in response to the optical density detected by a centrate optical density sensor, which corresponds to the level of cells in the centrate. For example, if the detected cell level in the centrate exceeds a set limit, the controller may control the centrate flow rate to decrease. This may be done by the controller separately or in conjunction with controlling the concentrate discharge flow rate. The controller may vary the centrate flow rate to ensure that the cell level in the centrate remains below a set limit or within a set range.

[0147] Alternatively, or in addition, the controller may control the flow rate of the cell suspension into the disposable structure by varying the flow rates of the clarifier and concentrate from the disposable structure to maintain cell levels in the clarifier and concentrate within programmed limits stored in memory associated with the controller. The controller may also operate in accordance with its programming to vary other process parameters, such as bowl rotation speed, diluent introduction, and diluent introduction rate, to maintain clarifier and concentrate characteristics that implement the set limits and desired process rate. In other exemplary configurations, other properties or parameters may also be monitored or adjusted by the control system to achieve the desired product.

[0148] 27 is a cross-sectional view of yet another disposable centrifuge structure 314. This centrifuge structure 314 is generally similar to the disposable separator structure 178, except that this disposable structure 314 includes an element extending therein that more reliably maintains the air / liquid interface of the air pocket separating the seal 236 from the liquid being processed in a desired radial position during operation.

[0149] In the disposable configuration 314, the separator pump 208 is located within a separator pump chamber 316. The separator pump chamber 316 is vertically bounded at its bottom by a lower separator centripetal pump chamber surface 318 and at its top by a circular upper separator centripetal pump chamber surface 320.

[0150] The lower centrate pumping chamber surface 318 extends radially outward from a lower centrate centripetal pumping chamber opening 322. In the exemplary configuration, the lower centrate centripetal pumping chamber opening 322 extends through the circular top of the core 200, corresponding to the top opening 202. The supply line 184 extends through the lower centrate centripetal pumping chamber opening.

[0151] An upper centrate centripetal pumping chamber surface 320 extends radially outward from a circular upper centrate centripetal pumping chamber opening 324. The supply line 184 and the centrate discharge line 212 extend axially through the upper centrate centripetal pumping chamber opening.

[0152] A plurality of angularly spaced, upwardly extending lower centrate chamber vanes 326 extend above the lower centrate centripetal pumping chamber surface 318. The lower centrate chamber vanes 326 originate at the lower centrate centripetal pumping chamber opening 322 and extend radially outward. The lower centrate chamber vanes 326, shown in more detail in FIG. 28, extend radially outward from the rotational axis 174 a distance V. In an exemplary configuration, the lower centrate chamber vanes 326 extend upward within a circular recess in the lower centrate centripetal pumping chamber surface 318. Note that this configuration is illustrative only, and other configurations may be used. For example, the radial length of the vanes, the vane height, and the recess depth and diameter may be varied to achieve desired fluid pressure characteristics.

[0153] A plurality of angularly spaced, downwardly extending upper centrate chamber vanes 328 extend from the upper centrate centripetal pumping chamber face 320. Each upper centrate chamber vane 328 originates at an upper centrate centripetal pumping chamber opening 324 and extends radially outward. The upper centrate chamber vanes extend radially outward from the rotational axis 174 a distance known as the upper centrate vane distance. In the exemplary configuration, the upper centrate vane distance corresponds approximately to the lower centrate vane distance V. In the exemplary configuration, the upper centrate chamber vanes extend downward into circular recesses in the upper centrate centripetal pumping chamber face with a configuration similar to the lower centrate chamber vanes shown in FIG. 28, but in the opposite direction.

[0154] In the exemplary configuration shown, the centrate centripetal pump 208 includes a generally annular centrate pump opening 330. The generally annular centrate pump opening 330 extends radially outward from the rotational axis 174 a distance that is the centrate pump opening distance. The centrate pump opening distance at which the centrate centripetal pump opening 330 is located is greater than the lower centrate vane distance and the upper centrate vane distance for reasons that will be explained below.

[0155] In the exemplary configuration of the disposable structure 314, the concentrate centripetal pump 216 is located within a concentrate pump chamber 332. The concentrate pump chamber 332 is vertically bounded on its lower side by a circular lower concentrate centripetal pump chamber surface 334. The centrate pump chamber 332 is vertically bounded on its upper side by a circular upper centrate centripetal pump chamber surface 336.

[0156] A lower concentrate centripetal pumping chamber surface 334 extends radially outward from a lower concentrate centripetal pumping chamber opening 338. In the exemplary configuration, the lower concentrate centripetal pumping chamber opening corresponds in size to and is continuous with the upper centrate centripetal pumping chamber opening 324. The supply line 184 and the centrate discharge line 212 extend through the lower concentrate centripetal pumping chamber opening 338.

[0157] A plurality of angularly spaced, upwardly extending lower concentrate chamber vanes 340 extend above the lower concentrate centripetal pumping chamber surface 334. The lower concentrate chamber vanes 334 originate at the lower concentrate centripetal pumping chamber opening 338 and extend radially outward. The lower concentrate chamber vanes 334 extend radially outward from the axis of rotation the distance of the lower concentrate vane. In an exemplary configuration, the lower concentrate centripetal pumping chamber vanes 334, like the upper and lower centrate chamber vanes, extend above a circular recess in the lower concentrate centripetal pumping chamber surface. Of course, this configuration is illustrative only.

[0158] An upper concentrate centripetal pumping chamber surface 336 extends radially outward from an upper concentrate centripetal pumping chamber opening 342. The supply line 184, the centrate discharge line 212, and the centrate discharge line 220 extend coaxially through the upper concentrate centripetal pumping chamber opening 342. A plurality of angularly spaced upper concentrate chamber vanes 344 extend downward from the surface 336. The upper concentrate chamber vanes extend radially outward from the upper concentrate centripetal pumping chamber opening 342 a distance equal to the upper concentrate vane distance. The upper concentrate chamber vanes extend within upwardly extending circular recesses in the upper concentrate centripetal pumping chamber surface. In an exemplary configuration, the upper concentrate chamber vanes are configured similarly to the lower concentrate chamber vanes and the upper and lower centrate chamber vanes described above. Of course, this approach is merely exemplary, and other approaches are possible in other configurations.

[0159] The concentrate centripetal pump 216 includes a generally annular concentrate pump opening 346. The concentrate pump opening is radially disposed from the axis of rotation 174 at a distance equal to the concentrate pump opening distance. Of course, this configuration is illustrative only, and other approaches are possible in other configurations.

[0160] In the exemplary disposable structure 314, the upper and lower concentrate chamber vanes 344, 340 and the upper and lower separate chamber vanes 326, 328 operate to stabilize and radially position the annular air / liquid interface 348 in the separate pump chamber 330 and the air / liquid interface 350 in the separate pump chamber 332. As shown in FIG. 28 , the air / liquid interface 348 is located radially midway along the length of the separate chamber vanes, i.e., radially inward from the separate pump opening 330. The radially extending separate chamber vanes provide a centrifugal pumping force to maintain the annular air / liquid interface 348 radially inward from the separate pump opening 330 both above and below the separate centripetal pump. In the exemplary configuration, the vanes further act to maintain the air / liquid interface, thereby maintaining a coaxial circular configuration both above and below the separate pump. The exemplary configuration further allows the radial position of the interface relative to the axis of rotation to be controlled as described below, thereby maintaining the centrate pump opening 330 in the centrate at all times and not exposing it to air.

[0161] The upper concentrate chamber vanes 344 and the lower concentrate chamber vanes 340 operate similarly to the separate chamber vanes. These vanes maintain a circular air / liquid interface 350 within the concentrate pump chamber 332 at a radial distance inside the generally annular concentrate pump opening 346. This configuration ensures that the concentrate pump opening is always exposed to the concentrate but not to the air. Note that in the illustrated configuration, the separate and concentrate centripetal pumps are approximately the same size; however, in other configurations, the sizes of the centripetal pumps may be different. In such cases, the separate and concentrate chamber vanes may extend at different radial distances from the axis of rotation. The air / liquid interfaces in the separate and concentrate pump chambers may also be located at different radial positions relative to the axis of rotation. Many different vane configurations are possible, depending on the specific relationship between the components comprising the disposable device and the specific material being processed by the disposable structure.

[0162] FIG. 30 illustrates the upper portion of yet another disposable structure 352. This disposable structure 352 is similar to the disposable structure 304, except as follows: The disposable structure 352 includes an air line 354 extending coaxially around the concentrate discharge line 220. The air line 354 communicates with openings 356 within the disposable structure. These openings 356 extend from the interior of the air line to above the concentrate centripetal pump 216 in the concentrate pump chamber 332. In this exemplary configuration, a seal 236 operatively engages the air line 354, maintaining airtight engagement with the air line as well as the concentrate discharge line, the centrate discharge line, and the supply line. The air line can be utilized to selectively maintain air pressure levels in air pockets within the disposable structure. This configuration is applicable to the above-described system and other systems that utilize an external source of pressurized air to separate the seals of the centrifuge structure from the material being processed while maintaining an air / liquid interface at a desired location. Of course, this structure is merely exemplary and other approaches may be employed in other configurations.

[0163] 31 is a schematic diagram illustrating a system 358 that can be used to continuously separate a cell suspension into a substantially cell-free centrate and concentrate. System 358 is similar to system 170 described above, with the following exceptions: In the illustrated configuration, system 358 operates using a disposable structure similar to disposable structure 352. Controller 274 of system 358 operates to control the position of the air / liquid interface within the disposable structure and ensures that the interface is maintained radially inward relative to the axes of rotation of the centrate pump opening and the concentrate pump opening, respectively.

[0164] In the exemplary configuration, the stream backpressure regulator 360 is fluidly connected to the centrate discharge line 262. In this exemplary configuration, the stream backpressure regulator 360 is fluidly intermediate the centrate discharge line 212 and the centrate pump 266. The exemplary system 358 includes a pressurized air source 362. The pressurized air source 362 is connected to a pressure control pilot valve 364. The control valve is operatively connected to a controller 274. In response to a signal from the controller 274, a variable pressure is selected in a pilot line 366. The pilot line 366 is fluidly connected to the backpressure regulator 360. The pressure applied by the pressure control pilot valve in the pilot line 366 can control the centrate flow, and therefore the centrate flow backpressure applied by the stream backpressure regulator 360.

[0165] In the exemplary configuration, pressure control valve 368 is in fluid communication with pressurized air source 362. Control valve 368 is also operatively connected to controller 274. In this exemplary configuration, control valve 368 is controlled to selectively apply precise pressures to air lines 354 and air pockets within the upper portion of disposable structure 352.

[0166] In the exemplary configuration, controller 274 operates according to stored executable instructions to control the operation of system 358 as described above in connection with system 170. In the exemplary configuration, controller 274 also operates to control pilot pressure valve 364 to vary the backpressure applied by backpressure regulator 360 to centrate discharge line 212. Controller 274 also controls valve 368. The controller operates to maintain and selectively vary the pressure applied to an upper air pocket within the disposable structure. The controller operates according to its programming to vary the centrate flow backpressure and / or air pocket pressure to maintain the air / liquid interface of the air pocket at an axis of rotation distance oriented inward from centrate pump opening 330 and concentrate pump opening 346. The pressure variations in both the centrate flow backpressure and the air pocket, in conjunction with the centrate chamber vanes and retentate vanes of this exemplary configuration, maintain the stability and radial outward extent of the air / liquid interface, thereby reliably limiting the introduction of air from the disposable structure into the centrate and retentate outputs. Furthermore, selectable centrate backpressure and flow can have a significant impact on the cell level of the discharged retentate and the corresponding detected optical density. Thus, the controller, operating according to its programming, can selectively vary the retentate flow rate, centrate backpressure, centrate flow rate, air pocket pressure, cell suspension flow rate to the disposable structure, and other possible operating variables of the centrifugation process, maintaining the centrate and retentate characteristics within set limits and / or set ranges stored in at least one storage device associated with the controller. Furthermore, the exemplary configuration allows for the separation and manipulation of different types of materials at different flow rates while maintaining reliable control of the separation process. Of course, although control of the position of the air / liquid interface has been described with respect to features of system 170, such control may also be employed in other types of systems having other or different types of processing elements.

[0167] 32-34 illustrate yet another disposable structure 370. This exemplary disposable structure 370 includes various features similar to those described in connection with the other disposable structures 178, 240, and 250. Note that additional features used in the other disposable structures and described herein may also be used in the features and related configurations illustrated in disposable structure 370.

[0168] The disposable structure 370 includes an upper disk-shaped portion 372. This exemplary upper disk-shaped portion 372 includes a separator centripetal pump chamber 374. The separator centripetal pump 208 is housed within the separator centripetal pump chamber 374. The separator centripetal pump chamber includes a separator chamber volume within the upper disk-shaped portion.

[0169] The separator centripetal pump chamber is in fluid communication with the separation chamber via at least one separator channel 410. The separator channel 410 is in fluid communication with at least one separator channel inlet 412. The exemplary at least one separator channel inlet 412 is in fluid communication with the separation chamber proximate to, but extending radially outward from, the cylindrical wall of the cylindrical core. In the exemplary configuration shown, the at least one separator channel inlet 412 is a substantially annular, unitary inlet, and the separation channel is a substantially annular, unitary channel.

[0170] Additionally, the upper disk-shaped portion includes a concentrate centripetal pumping chamber 376. The exemplary concentrate pumping chamber 376 is a cylindrical chamber that is horizontally partitioned by a vertically extending circular partition wall 378. The concentrate centripetal pumping chamber 376 includes a concentrate chamber volume within the upper disk-shaped portion. In this exemplary configuration, the volume of the centrate chamber within the upper disk-shaped portion is larger than the volume of the concentrate chamber for reasons described below.

[0171] The exemplary upper disk-shaped portion includes an upper piece 394 and a lower piece 396. In the exemplary configuration, the upper and lower pieces are held together by a releasable engagement. This method is exemplary, and other methods may be used in other configurations. In an operational or operating state, the exemplary lower piece 396 abuts on its upper side against an upper annular boundary surface 398. The exemplary lower piece 396 abuts on its lower side against a lower annular boundary surface 400. The upper annular boundary surface 398 has a radially outwardly facing conical annular upper surface portion 402 and a radially inwardly facing, radially planar extending upper surface portion 404. The lower annular boundary surface 398 has a radially outwardly facing, conical annular lower surface portion 406 and a radially inwardly facing, radially planar extending lower surface portion 408. In the operative position of upper piece 394 and lower piece 396, radially inwardly facing, radially planar, horizontally extending lower surface 408 and radially inwardly facing, radially planar, horizontally extending upper surface 404 extend parallel to one another, although in the operative position conical annular upper surface portion 402 and conical annular lower surface portion need not be parallel for reasons that will be explained below.

[0172] In the exemplary configuration, a substantially annular cell concentrate channel 380 extends between the upper piece 394 and the lower piece 396 of the upper disk-shaped portion 372. The annular cell concentrate channel 380 extends radially inward from a substantially annular cell concentrate channel inlet 382. The concentrate channel inlet is located further radially outward from the separation chamber inlet 412. The concentrate channel inlet 382 is located in fluid communication with an upper region of the separation chamber 224 at the radial periphery adjacent the inner surface of the outer wall where the cell concentrate 384 collects in the annular radially outward region 384 of the separation chamber upon rotation of the device by the centrifuge bowl, as shown in FIG.

[0173] In the exemplary configuration, a substantially annular funnel channel 381 extends upward and radially inward relative to an annular cell concentrate channel inlet 382. In the operating position, an exemplary lower piece 396 of the upper disk-shaped portion 372 abuts a substantially planar, radially extending surface 379 on its radially lower, inward-facing side. The exemplary radially extending surface 379 terminates radially outward at an annular edge 377. In the operating position of the disposable configuration, the annular funnel channel 381 extends outward from the annular edge 377. Additionally, an exemplary upper piece 394 of the exemplary annular, disk-shaped portion 372 has a substantially annular cell concentrate guide surface 383. The annular cell concentrate guide surface 383 extends below the annular funnel channel 381 and abuts the separation chamber 224 radially outward at the axial level of the radially extending, planar surface 379. In this exemplary configuration, the annular cell concentrate guide surface 383 further extends radially outward and upwardly adjacent the funnel channel.

[0174] In this exemplary configuration, the annular cell concentrate channel 380 terminates radially inwardly in the concentrate centripetal pump chamber 376 at a substantially annular cell concentrate outlet 386. Also in the exemplary configuration, the annular cell concentrate outlet 386 is located at a position extending to the midpoint of the vertically extending boundary wall 378. In this exemplary configuration, the concentrate centripetal pump has a substantially annular concentrate centripetal pump inlet 388. The annular cell concentrate outlet of the channel 380 is radially and axially aligned with the concentrate centripetal pump inlet 388.

[0175] In the exemplary configuration, the annular cell concentrate channel has a tapered portion 390 and a radially extending portion 392 in an axial cross-section. The radially extending portion 392 extends radially outward directly between the radially planar upper surface portion 404 of the bottom piece 396 and the radially planar lower surface portion 408 of the top piece 394. Furthermore, the exemplary radially extending portion 392 extends radially outward directly from the annular cell concentrate outlet 386. In the disposable configuration's operating position, the horizontally radially extending portion 392 of the annular cell concentrate channel has a constant channel height. That is, the channel height refers to the dimension of the channel transverse to the direction of concentrate flow in each region of the channel. As a result, the horizontally radially extending portion has a constant cross-sectional area throughout its entire length. In this exemplary configuration, the horizontally and radially extending portions 392 are axially and radially aligned, and therefore level in axial cross section with the inlet of the concentrate centripetal pump.

[0176] The tapered section 390 of the annular cell concentrate channel 380 extends between the conical annular upper surface section 402 of the bottom piece and the conical annular lower surface section 406 of the top piece. The annular cell concentrate channel 380 is configured with a channel portion in which the channel height (and therefore the channel cross-sectional area) increases continuously from the cell concentrate flow inlet 382 to the location where the tapered section fluidly connects to the radially extending section 392. This configuration increases the cross-sectional area of ​​the channel portion 390 perpendicular to the direction of concentrate flow and increases its proximity to the axis of rotation. The continuously increasing cross-sectional area of ​​the concentrate channel means that the channel cross-sectional area perpendicular to the direction of concentrate flow increases smoothly throughout the channel portion, without any discrete steps in locations where the channel cross-sectional area changes by more than 10%.

[0177] In this configuration, the cell concentrate fluid velocity can be maintained at a desired high velocity due to the constant increase in channel height close to the axis of rotation of the disposable system within tapered section 390. In the exemplary configuration, the annular cell concentrate channel avoids pressure drop areas, thereby maintaining a desired radial inward velocity of the cell concentrate components from channel inlet 382 to channel outlet 386.

[0178] In exemplary configurations, the annular inlet 382 to the annular channel 380 is the portion of the channel with the smallest height and smallest cross-sectional area in the axial cross-section. At the annular inlet 382, ​​cells and the fluid in which they are suspended begin to flow radially inward. In this region of the disposable structure, the cells, being denser than the liquid, experience centrifugal acceleration forces that are directed radially outward by the rotation of the centrifuge. During operation of the exemplary configurations, an external concentrate pump, such as the concentrate pump 272 described above, is activated to maintain a flow rate that results in an average velocity of the radially inward liquid flow at the annular channel inlet 382. This maintains the settling velocity and the radially outward force acting on the cells resulting from the centrifugal forces acting on the concentrate at the channel inlet. Furthermore, in some exemplary configurations, the upwardly tapered configuration of the tapered section 390 ensures that the component of the sedimentation force opposing cell flow at the channel inlet and channel section is smaller than the component of the sedimentation force opposing cell movement within the radially inward channel at the same radial location.

[0179] In an exemplary configuration, the height of the annular tapered portion of the annular concentrate channel increases with decreasing radial distance from the axis of rotation of the disposable structure. In an exemplary configuration of the channel in an axial cross-section, the channel height of the tapered portion 390, and therefore the cross-sectional area perpendicular to the direction of concentrate flow, gradually increases with increasing proximity to the axis of rotation of the disposable structure (decreasing radial distance from the axis of rotation). The exemplary configuration in which the channel height gradually increases with decreasing radial distance from the axis is preferred because cells in the cell concentrate, along with the liquid in which they are suspended, continuously pass through the concentrate channel at a high radial inward velocity. Despite the increased channel height, the cells are subjected to a reduced radial outward centrifugal acceleration force and the corresponding decrease in radial distance from the axis of rotation allows the cell concentrate to maintain a preferred radial inward velocity throughout its radial inward transition from the channel inlet 382. Thus, in the exemplary configuration, as the cell concentrate moves radially inward from the annular inlet 382 toward the cell concentrate outlet 386 and into and through the concentrate centripetal pump chamber 376, the cell concentrate maintains a suitably high radial inward velocity within channel portion 390 throughout the entire channel 380.

[0180] It should be noted that while the exemplary configurations shown in Figures 32-34 feature a channel portion with a continuously increasing cross-sectional area beginning at the channel inlet, other configurations may employ other approaches or configurations. For example, in some other configurations, channel portions with this configuration may be located at other locations. Such locations may depend on the specific configuration of the concentrate channel and the conditions necessary for the concentrate and cells therein to move in a manner that achieves a sufficiently high flow velocity through the specific portion of the channel, thereby suppressing sedimentation and other forces that would impede the intended flow. Furthermore, while this exemplary configuration uses a single annular concentrate channel portion with a continuously increasing cross-sectional area, other configurations may use other channels, such as multiple concentrate channels. The configurations shown in Figures 32-34 are exemplary only, and other configurations may be used.

[0181] In an exemplary configuration, the concentrate outlet line 220 is fluidly connected to an external concentrate pump during system operation, as previously described. In an exemplary configuration, the concentrate pump can be operated similarly to system 170 previously described, or similarly to other systems responsive to control circuitry that generates an outlet flow of cell concentrate while maintaining a suitable backpressure in the concentrate outlet line. Of course, many other components can be implemented in a system that operates the disposable structure 370 to provide the operational capabilities listed herein.

[0182] During operation of the exemplary system, the disposable structure 370 connects to a centrifuge bowl and separates the cell suspension in the structure's interior region. The cell suspension is separated into a substantially cell-free cell separation solution and a cell-enriched cell concentrate solution, as previously described. In the exemplary configuration, centrifugal action creates an annular cell concentrate region 384 at the radial periphery of the upper separation chamber. This exemplary annular cell concentrate region maintains fluid communication with the annular channel inlet 382, ​​as well as contact with the substantially annular cell concentrate guiding surface and the annular funnel channel along which the cell concentrate moves toward the channel inlet 382.

[0183] In some exemplary configurations, an annular cell concentrate guiding surface 383 that borders the radial periphery of the separation chamber adjacent the radially extending surface 379 of the upper disk-shaped portion acts to urge the cell concentrate upward, forcing it upward into the annular funnel channel 381. This is because the guiding surface extends further radially outward and upwardly closer to the final flow path. Centrifugal forces generated by the rotation of the centrifuge move the cell concentrate radially outward against the annular cell concentrate guiding surface 383, allowing it to engage the guiding surface and enter the annular funnel channel. The guiding surface guides the cell concentrate, causing it to move upward and radially inward into the annular cell concentrate inlet. As the cell concentrate is guided radially upward toward the channel inlet 382 by the annular converging surface whose axial cross-section borders the annular funnel channel, the radial inward velocity of the liquid component of the cell concentrate increases as the area of ​​the funnel channel decreases. As previously discussed, the exemplary configuration has minimal height (and therefore minimal cross-sectional area), thereby maximizing the fluid velocity of the liquid phase of the cell concentrate at the annular channel inlet 382.

[0184] In the exemplary configuration and method of operation, an external concentrate pump operates to generate a flow of cell concentrate within the annular cell concentrate channel 380 from the annular cell concentrate inlet 382 to the annular cell concentrate outlet 386 at a flow rate that produces an average radially inward velocity of the cell concentrate liquid phase that is greater than the settling velocity of the cells. The exemplary configuration achieves a constant, high average velocity of the cell concentrate liquid phase across the height of the channel and throughout the entire radial length of the channel, thereby favoring flow of the cell concentrate and the cells therein into the annular cell concentrate channel. Furthermore, in the exemplary configuration, due to the continuously increasing cross-section of the tapered portion 390 of the channel, the constant height of the radially extending portion 392, and the generally constant, relatively small cross-sectional area of ​​the generally annular cell concentrate channel, there are no regions of significant pressure drop along the length of the channel, and liquid velocity and cell continuity throughout the channel are adequately maintained.

[0185] Furthermore, during operation of the exemplary configuration, the cell concentrate entering the concentrate centripetal pump chamber 376 moves at a velocity and flow rate high enough to ensure that the liquid and cell phases of the cell concentrate pass radially inward through the concentrate centripetal pump inlet. In this exemplary configuration, this result is facilitated not only by the volume and location of the concentrate centripetal pump chamber, but also by the location of the channel outlet 386 relative to the centripetal pump inlet 388 of the centripetal pump 216.

[0186] These features of the exemplary configuration facilitate radial inward flow of cell concentrate in the exemplary disposable structure and provide advantageous operation of the disposable structure and corresponding system. These features and configurations are exemplary, and the principles described herein can be applied to other configurations and other disposable or multi-use structures to achieve desired performance characteristics and cell separation in other centrifugation processes.

[0187] 35-38 illustrate yet another alternative configuration for the disposable structure 414. This exemplary alternative disposable structure has many of the features already described and includes an upper disk-shaped portion 416. An outer wall 418 is configured to operatively connect with the centrifuge bowl in which the disposable structure 414 is located. The structure includes a lower portion 420 of the outer wall 418. The disposable structure illustrated in FIG. 36 has an interior region that is frusto-conical in shape with a smaller inner diameter adjacent the lower portion 420.

[0188] Similar to previously described configurations, the disposable structure 414 has a cylindrical core 422 that extends axially between upper and lower portions of the structure's interior region and has a cylindrical outer partition wall 424. Note that while the cylindrical core 422 is shown as a solid structure in FIG. 36, hollow core structures can be used in other configurations. In the exemplary configuration, the cylindrical core 422 extends within the interior region between the bottom of the upper disk-shaped portion and a plurality of upwardly directed, angularly spaced vanes 426. The vanes 426 have fluid channels between them that extend upwardly from the interior of the wall that borders the lower portion of the disposable structure's interior region.

[0189] In the exemplary configuration, the disposable structure 414 is configured to rotate about an axis 428 within the centrifuge bowl. Additionally, the disposable structure has a vertically extending supply tubing 430 in the operating position that receives the cell culture medium into an interior region of the structure. The exemplary supply tubing 430 extends downward to a tubing opening 432 in the lower portion of the disposable structure to an area where the influent to be separated into a cell separation solution and a cell concentrate solution is introduced. In the exemplary configuration shown, the supply tubing 430 extends axially through a cylindrical opening 434 in the core.

[0190] In its operating position, the exemplary disposable structure includes a vertically extending separate solution discharge line 436. The vertically extending separate solution discharge line 436 is fluidly connected to a separate solution centripetal pump 438. The separate solution centripetal pump is located within a separate solution centripetal pump chamber 440 located within the upper disk-shaped portion 416. The separate solution centripetal pump chamber 440 is fluidly connected to a separation chamber 442, which extends radially between the outer wall of the core 422 and a wall bounding the interior region of the disposable structure. The separate solution centripetal pump chamber is fluidly connected to the separation chamber 442 via at least one separate solution channel inlet 444 and at least one separate solution channel 446. In the exemplary configuration, the at least one separate solution channel inlet 444 is located in the separation chamber radially adjacent to, but radially outward from, the cylindrical wall bounding the core 422. In the exemplary configuration, the separation solution channel inlet is arcuate in shape. Furthermore, in the operating position of the exemplary configuration, the centrate centripetal pumping chamber 440 has horizontally extending upper and lower centrate pumping chamber surfaces which may have upper and lower centrate chamber vanes such as vanes 326 and 328 previously described.

[0191] Additionally, the exemplary disposable structure 414 includes a concentrate discharge line 448 extending perpendicular to the operating position. As with the other configurations described above, the concentrate discharge line 448, the centrate discharge line 436, and the supply line 430 are coaxially arranged on the disposable structure. The concentrate discharge line 448 is fluidly connected to a concentrate centripetal pump 450, which is located within a concentrate centripetal pump chamber 452 within the upper disk-shaped portion 416. In the exemplary configuration, the concentrate centripetal pump chamber is bounded by upper and lower concentrate centripetal pump chamber surfaces, each of which may have radially extending chamber vanes similar to those previously described.

[0192] In this exemplary configuration, the concentrate centripetal pumping chamber is fluidly connected to the separation chamber via a plurality of radially extending concentrate channels 454. In the exemplary configuration, each concentrate channel extends within the upper disk-shaped portion and is angularly spaced apart from each other channel. The exemplary upper disk-shaped portion 416 comprises an upper piece 456 and a lower piece 458. The exemplary upper disk-shaped portion further includes a bottom piece 460. In the exemplary active configuration, the upper piece 456, the lower piece 458, and the bottom piece 460 are in sandwich-like engagement. In the active position, each of the plurality of concentrate channels abuts at least one downward-facing surface of the upper piece 456 and at least one upward-facing surface of the lower piece 458.

[0193] As shown in FIG. 36 , each of the plurality of concentrate channels 454 has a concentrate channel inlet 462. In an exemplary configuration, the concentrate channel inlet has the smallest cross-sectional area perpendicular to the direction of concentrate flow throughout each concentrate channel. Each concentrate channel inlet 462 fluidly connects to the periphery of the separation chamber through a vertical opening 464 that extends into the upper disk-shaped portion. In an exemplary configuration, each concentrate channel 454 fluidly connects to the separation chamber through a respective vertical opening 464 that extends through the bottom piece 460 and is located between and borders the upper and lower pieces 456, 458. In an exemplary configuration, each vertical opening 464 comprises an arc-shaped, elongated slot in the bottom piece 460. It will be appreciated that this arrangement is exemplary and that other approaches may be used in other configurations.

[0194] In an exemplary configuration, each concentrate channel has a generally constant cross-sectional width from the channel inlet 462 to each opening of the channel into the concentrate centripetal pumping chamber. Each concentrate channel has a channel portion with a continuously increasing cross-sectional area perpendicular to the direction of concentrate flow within each concentrate channel portion. This cross-sectional area increases with increasing proximity of the channel portion to the axis of the disposable structure. In an exemplary configuration, the multiple concentrate channels are configured such that the cross-sectional area perpendicular to the direction of concentrate flow increases continuously at the tapered portion 468 as a result of varying channel heights in the tapered portion. The height of each channel increases with increasing proximity to the axis of rotation. Of course, this configuration is exemplary, and other approaches can be used in other configurations.

[0195] In the exemplary configuration, each channel portion of successively increasing cross-sectional area begins at a respective concentrate channel inlet 462 and continues through an upwardly and radially inwardly extending tapered portion 468. Each tapered portion 468 fluidly connects to a horizontally and radially extending portion 470 of a respective concentrate channel 454. In the exemplary configuration, each horizontally and radially extending portion of each concentrate channel extends from the tapered portion 468 to the concentrate centripetal pumping chamber 450. In the exemplary configuration, the horizontally and radially extending portion of the concentrate channel has a constant cross-sectional area perpendicular to the concentrate flow from the tapered portion radially inward to the concentrate centripetal pumping chamber. Note that this configuration is exemplary, and other approaches may be applied in other configurations.

[0196] During operation of the exemplary disposable structure 414, the upper disk-shaped portion 416, wall 418, and core rotate and connect to the centrifuge bowl. The supply line 430, the centrate discharge line 436, and the concentrate discharge line 448 remain stationary, along with the concentrate centripetal pump 450 and the centrifugal pump 438. In the exemplary configuration, the cell culture separates into a cell detachment solution and a cell concentrate in the separation chamber 442 by rotation-induced centrifugal forces. The cell concentrate collects in the upper and radially outer region of the separation chamber 442, while the substantially cell-free detachment solution collects in the region of the separation chamber adjacent the outer cylindrical wall of the core.

[0197] Centrate fluid enters the separation chamber via multiple separation channel inlets 444 and exits the chamber via the separation centripetal pump 438 and separation outlet piping. An external concentrate pump, such as the concentrate pump previously described, is operatively connected to the concentrate outlet piping, causing the concentrate to flow out of the disposable separation chamber. This concentrate flow causes the cell concentrate to move upward through the vertical openings 464 and through the concentrate inlets 462, where the relatively small cross-sectional areas of the concentrate channel inlets result in high velocity of the cell concentrate. The high fluid and cell flow velocity of the cell concentrate exerts a force on the cells contained within the cell concentrate, overcoming the radially outward force acting on the cells, as described in connection with the prior art configuration, causing the cells to move upwardly and radially inward into the tapered section 468.

[0198] The upwardly and radially inwardly extending tapered sections have a continuously increasing cross-sectional area perpendicular to the direction of concentrate flow. This continuously increasing cross-sectional area maintains a sufficiently high concentrate flow velocity at each radial location throughout the channel sections, resulting in the cell concentrate flowing into the concentrate chamber at a desired high velocity despite radially outward forces acting at each location in the channel sections. In the exemplary configuration, the cell concentrate exits the tapered sections of the concentrate channels and passes through the horizontally and radially extending sections of each channel to the concentrate centripetal pumping chamber 452, where the cell concentrate maintains a desired high velocity. As a result, in the exemplary configuration, the cell concentrate has flow characteristics that promote cell concentrate flow within the disposable structure and facilitate the cell separation process. Of course, the configuration of the disposable structure 414 is exemplary, and other configurations may utilize other configurations.

[0199] In some exemplary configurations, the top, bottom, and bottom pieces of the upper disk-like portion 416 may be releasably engaged. This facilitates manufacturing and allows for inspection, cleaning, etc. In other exemplary configurations, the pieces may be permanently engaged. In still other exemplary configurations, structures similar to those described herein may be formed from other components that provide the useful properties and performance described herein. Furthermore, the configuration of the disposable structure 414 is exemplary, and the useful principles and structures described herein may be used in other separator structure configurations.

[0200] Thus, the novel centrifuge system and method, as illustratively disclosed, achieves at least some of the objectives set forth above, overcomes problems and overcomes challenges encountered when using conventional devices and systems, and achieves the desired results described above.

[0201] Certain terms have been used in the foregoing description for the purposes of brevity, clarity, and ease of understanding, but without unnecessary limitation. These terms are intended to be descriptive and broad. Moreover, the descriptions and illustrations herein are exemplary, and the invention is not limited to the exact details shown and described.

[0202] It should be noted that features and / or relationships associated with one exemplary configuration may be combined with features and / or relationships in other exemplary configurations, i.e., various features and / or relationships may be combined in yet other configurations. The inventive scope of the present disclosure is not limited to the exemplary configurations illustrated herein and shown in the drawings.

[0203] In the claims, features recited as means for performing a function should be construed to encompass any means known to one of ordinary skill in the art to perform that function, and are not limited to the structures shown in this specification or mere equivalents thereof.

[0204] The discoveries and principles of new and useful features have been described, and the manner of making, utilizing and operating them, and the advantages obtained, new and useful structures, apparatus, elements, arrangements, parts, combinations, systems, acts, methods and relationships are set forth in the appended claims. [Explanation of symbols]

[0205] 10 Core 12 axis 14 Stationary device 16 Equipment 18 Supply piping 20 aperture 22 Cavity 24 Separation liquid centripetal pump 26 Entrance opening 28 Exit opening 30 Separated liquid piping 32 Separation liquid pump room 34 Concentrate pump 36 Entrance opening 38 Exit opening 40 Concentrate outlet piping 42 Concentrate pump room 44 Upper 46 Fluid seal 48 Accelerator Blade 50 Tapered Wall 52 Concentrate Slot 54 Circular Wall 56 Separation liquid hole 58 Plate 1 60 Second Plate 62 screws 64 Spiral Channel 66 Outer circumference 67 Exterior Wall 68 Entrance opening 70 Circular Recovery Room 74 Wall 76 Wall 78 Wall 80 circular surfaces 82 Bowl 84 axis 86 Motor 88 Core 90 Cavity / Separation Chamber 92 Suspension inlet supply piping 94 Entrance opening 96 Centripetal Pump 98 Entrance 100 Pump outlet 102 Separated liquid outlet piping 104 Upper 106 Stickers 108 Pump 110 Entrance Line 112 Concentrate Line 114, 116 valves 118 Separation liquid discharge line 120 Separation liquid discharge pump 122 Pressure Damping Reservoir 124 bottom port 126 Upper Port 128 Upper Port 130 Sterilizing Filter 132 Regulator 134, 136 and 138 Liquid Level Sensors 140 valves 142 control circuit 143 Air Line 144 processors 145 Aperture 146 Data storage devices 147 Liquid level 148 Subroutine Processes 150 processes 152 process 154 Process 156 process 158 Process 160 processes 162 Process 170 Centrifuge System 172 Centrifuge Bowl 174 axis 176 Cavity 178 Disposable structure 180 Top opening 182 Fixed structure 184 Supply piping 186 Internal area 188 1st shaft end 190 Aperture 192 2nd shaft end 194 disk-shaped part 196 Circular Outer 198 Circular Boundary Wall 200 Core 202 Upper opening 204 Lower opening 206 Separation liquid centripetal pump chamber 208 Separation liquid centripetal pump 210 Separation liquid opening 212 Separated liquid discharge piping 214 Concentrate pump chamber 216 Concentrate pump 218 Concentrate opening 220 Concentrate discharge piping 222 Exterior Wall 224 Separation room 226 Textured exterior 228 dent 230 Lower opening 232 Disk-shaped part 234 Channel 236 Seal 238 Centrifuge System 240 Disposable structure 242 Lower disk-shaped part 244 flow path 246 Feather 248 Centrifuge System 250 Disposable structure 252 Lower disk-shaped part 254 Accelerator Blade 256 motor 258 Cell Culture Supply Line 260 Supply Pump 262 Separation liquid discharge line 264 Separation Liquid Optical Density Sensor 266 Separation Pump 268 Concentrate discharge line 270 Concentrate Optical Density Sensor 272 Concentrate Pump 274 Control Circuit 276 processors 280 process 284 Process 286 Process 288 Process 290 process 292 Process 294 Process 296 Process 298 Process 300 processes 302 process 304 Disposable structure 306 Annular concentrate dam 308 Tapered outer surface 310 Separation Dam 312 Annular recess 314 Disposable Centrifuge Structure 316 Separation liquid pump room 318 Lower separated liquid centripetal pump chamber surface 320 Upper separated liquid centripetal pump chamber surface 322 Lower separated liquid centripetal pump chamber opening 324 Upper separated liquid centripetal pump chamber opening 326 Lower separation liquid chamber blade 328 Upper separation liquid chamber blade 330 Separation liquid centripetal pump opening 332 Concentrate Pump Room 334 Lower concentrate centripetal pump chamber surface 336 Upper separated liquid centripetal pump chamber surface 338 Lower concentrate centripetal pump chamber opening 340 Lower concentrated liquid chamber blade 342 Upper concentrate centripetal pump chamber opening 344 Upper concentrated liquid chamber blade 346 Concentrate Pump Opening 348 Air / Liquid Interface 350 Air / liquid interface 352 Disposable Structure 354 Air piping 356 Aperture 358 System 360 Back pressure regulator 362 Pressurized Air Source 364 Pressure Control Pilot Valve 366 Pilot Line 368 Pressure Control Valve 370 Disposable structure 372 Upper disc-shaped part 374 Separation liquid centripetal pump chamber 376 Concentrate pump chamber 377 Circular Edge 378 Circular Partition Wall 379 Surface 380 cell concentrate channels 381 Funnel Channel 382 Cell concentrate channel entrance 383 Cell Concentrate Guideway 384 Cell concentrate 386 Cell concentrate outlet 388 Concentrate centripetal pump inlet 390 Tapered part 392 Radial extending portion 394 Upper Piece 396 Lower Piece 398 Upper annular boundary surface 400 Lower annular boundary surface 402 Conical annular upper surface 404 Top part 406 Conical annular lower surface part 408 Bottom part 410 Separation Channel 412 Entrance 414 Disposable Structure 416 Upper disk-shaped part 418 Exterior Wall 420 Lower part 422 Cylindrical Core 424 Exterior Partition Wall 426 Feather 428 axis 430 Supply piping 432 Pipe opening 434 Cylindrical opening 436 Separated liquid discharge piping 438 Separation liquid centripetal pump 440 Separation liquid centripetal pump chamber 442 Separation room 444 Separation channel inlet 446 Separation Channel 448 Condensate discharge piping 450 Concentrate Pump 452 Concentrate pump chamber 454 Concentrate Channel 456 Upper piece 458 Lower Piece 460 bottom piece 462 Concentrate channel inlet 464 Vertical opening 468 Tapered section 470 parts 1000 Centrifuge Structure 1100 Flexible Liner 1110 Thermal bonding part 1200 Lower flange 1210 Top 1300 Upper flange 1310 Bottom surface 1400 Centripetal Pump 1410 Pairing Disk 1415 Gap 1420 Rotary Pump Room 1420 holes 1500 Core Structure 1505 Top surface 1510 Core 1515 Exterior 1520 central cavity 1525 Rotating Axis 1530 Slit / hole 1540 hole 1550 Separation room 1555 Separation reservoir 1560 Accelerator Blade 1580 Plate 1590 Accelerator Bowl 1600 Centrifuge Cover 1605, 1620 Interior 1610 Engagement cap part 1630 Radial fin 1640 wall 1700 Rotating Mechanical Seal 2000 Supply / Discharge Equipment 2100 Supply piping 2110 Nozzle 2200 Discharge of separated liquid 2300 Supply Pipe Carrier 2400 Separated liquid outlet 2500 Discharge of concentrated liquid 2500 cell discharge tube 2510 Peristaltic Pump 3000 versatile structure 3100 Bowl 3200 Multi-Purpose Bowl Cover 3210 Navin 4400 Centripetal Pump 4410 Pairing Disk 4415 Gap 4420 Pump Room 4430 Concentration Sensor 4540 Slit / hole 4630 Accelerator Fin 5000 Dilution Solution Supply 5100 Disk-shaped flow channel 5150 Diluent Pump 6000 distance 6010 depth 6100 throttle tube R arrow V distance

Claims

1. 1. An apparatus having a structure releasably received within a rotatable centrifuge bowl, the structure disposed within the bowl operatively separating cells in a cell culture into a cell concentrate and a cell separation solution within an interior region of the structure, the apparatus comprising: In the operating position, this structure an upper disk-shaped portion; The bottom and a cylindrical core vertically intermediate said upper disk-shaped portion and said lower portion; a separation chamber disposed radially outward from and surrounding the core; an outer wall configured to operatively engage the bowl, extending in fluid-tight relation to said upper disk-shaped portion and bounding said separation chamber; extending in a surrounding relationship to the core and the separation chamber; an outer wall having a frusto-conical interior shape, the inner radius of the outer wall being smaller adjacent the lower portion than the inner radius of the outer wall adjacent the upper disk-shaped portion; a vertically extending supply pipe; a separated liquid discharge pipe extending vertically; a concentrated liquid discharge pipe extending vertically; and the upper disk-shaped portion and the outer wall are rotatable about a vertical axis while operatively engaging the bowl; the structure further includes a centrifugal pump axially aligned with the core, coaxially positioned about the supply line, and in fluid communication with the centrate discharge line; the separate liquid centripetal pump is located within a separate liquid centripetal pump chamber within the upper disk-shaped portion; the separator centripetal pumping chamber is in fluid communication with the separator chamber via at least one separator channel extending within the upper disk-shaped portion; each separation channel extends in fluid communication between a separation channel inlet located radially outward of the core and the separation centripetal pumping chamber; the structure further includes a concentrate centripetal pump axially aligned with the core, coaxially positioned about the supply line, positioned vertically above the centrate centripetal pump, and in fluid communication with the concentrate discharge line; the concentrate centripetal pump is located within a concentrate centripetal pump chamber within the upper disk-shaped portion; the concentrate centripetal pumping chamber is in fluid communication with the separation chamber via a plurality of radially extending concentrate channels extending within the upper disk-shaped portion; the plurality of radially extending concentrate channels extend radially between respective concentrate channel inlets located radially outward of each and every one of the separate liquid inlets; When the bowl rotates, the upper disk-shaped portion and the outer wall rotate relative to the supply pipe, the separated liquid discharge pipe, the concentrated liquid discharge pipe, the separated liquid centripetal pump, and the concentrated liquid centripetal pump, respectively; the plurality of concentrate channels have channel portions intermediate their respective concentrate channel inlets and the concentrate centripetal pumping chamber, the cross-sectional area of ​​which, perpendicular to the direction of concentrate flow through the channel portions, increases continuously as the cross-sectional area asymptotically approaches the perpendicular axis; and The device wherein said plurality of radially extending concentrate channels comprises a plurality of angularly spaced individual concentrate channels.

2. 1. An apparatus having a structure releasably received within a rotatable centrifuge bowl, the structure disposed within the bowl operatively separating cells in a cell culture into a cell concentrate and a cell separation solution within an interior region of the structure, the apparatus comprising: In the operating position, this structure an upper disk-shaped portion; The bottom and a cylindrical core vertically intermediate said upper disk-shaped portion and said lower portion; a separation chamber disposed radially outward from and surrounding the core; an outer wall configured to operatively engage the bowl, extending in fluid-tight relation to said upper disk-shaped portion and bounding said separation chamber; extending in a surrounding relationship to the core and the separation chamber; an outer wall having a frusto-conical interior shape, the inner radius of the outer wall being smaller adjacent the lower portion than the inner radius of the outer wall adjacent the upper disk-shaped portion; a vertically extending supply pipe; a separated liquid discharge pipe extending vertically; a concentrated liquid discharge pipe extending vertically; and the upper disk-shaped portion and the outer wall are rotatable about a vertical axis while operatively engaging the bowl; the structure further includes a centrifugal pump axially aligned with the core, coaxially positioned about the supply line, and in fluid communication with the centrate discharge line; the separate liquid centripetal pump is located within a separate liquid centripetal pump chamber within the upper disk-shaped portion; the separator centripetal pumping chamber is in fluid communication with the separator chamber via at least one separator channel extending within the upper disk-shaped portion; each separation channel extends in fluid communication between a separation channel inlet located radially outward of the core and the separation centripetal pumping chamber; the structure further includes a concentrate centripetal pump axially aligned with the core, coaxially positioned about the supply line, positioned vertically above the centrate centripetal pump, and in fluid communication with the concentrate discharge line; the concentrate centripetal pump is located within a concentrate centripetal pump chamber within the upper disk-shaped portion; the concentrate centripetal pumping chamber is in fluid communication with the separation chamber via a plurality of radially extending concentrate channels extending within the upper disk-shaped portion; the plurality of radially extending concentrate channels extend radially between respective concentrate channel inlets located radially outward of each and every one of the separate liquid inlets; When the bowl rotates, the upper disk-shaped portion and the outer wall rotate relative to the supply pipe, the separated liquid discharge pipe, the concentrated liquid discharge pipe, the separated liquid centripetal pump, and the concentrated liquid centripetal pump, respectively; the plurality of concentrate channels have channel portions intermediate their respective concentrate channel inlets and the concentrate centripetal pumping chamber, the cross-sectional area of ​​which, perpendicular to the direction of concentrate flow through the channel portions, increases continuously as the cross-sectional area asymptotically approaches the perpendicular axis; and the plurality of radially extending concentrate channels comprising a plurality of angularly spaced individual concentrate channels; the channel portions of the plurality of concentrate channels beginning at each concentrate channel inlet and extending radially inward and upward from said concentrate channel inlet; a constant cross-sectional width in a direction perpendicular to the direction of concentrate flow, and a vertical height that varies with radial distance from said axis; An apparatus comprising:

3. 1. An apparatus having a structure releasably received within a rotatable centrifuge bowl, the structure disposed within the bowl operatively separating cells in a cell culture into a cell concentrate and a cell separation solution within an interior region of the structure, the apparatus comprising: In the operating position, this structure an upper disk-shaped portion; The bottom and a cylindrical core vertically intermediate said upper disk-shaped portion and said lower portion; a separation chamber disposed radially outward from and surrounding the core; an outer wall configured to operatively engage the bowl, extending in fluid-tight relation to said upper disk-shaped portion and bounding said separation chamber; extending in a surrounding relationship to the core and the separation chamber; an outer wall having a frusto-conical interior shape, the inner radius of the outer wall being smaller adjacent the lower portion than the inner radius of the outer wall adjacent the upper disk-shaped portion; a vertically extending supply pipe; a separated liquid discharge pipe extending vertically; a concentrated liquid discharge pipe extending vertically; and the upper disk-shaped portion and the outer wall are rotatable about a vertical axis while operatively engaging the bowl; the structure further includes a centrifugal pump axially aligned with the core, coaxially positioned about the supply line, and in fluid communication with the centrate discharge line; the separate liquid centripetal pump is located within a separate liquid centripetal pump chamber within the upper disk-shaped portion; the separator centripetal pumping chamber is in fluid communication with the separator chamber via at least one separator channel extending within the upper disk-shaped portion; each separation channel extends in fluid communication between a separation channel inlet located radially outward of the core and the separation centripetal pumping chamber; the structure further includes a concentrate centripetal pump axially aligned with the core, coaxially positioned about the supply line, positioned vertically above the centrate centripetal pump, and in fluid communication with the concentrate discharge line; the concentrate centripetal pump is located within a concentrate centripetal pump chamber within the upper disk-shaped portion; the concentrate centripetal pumping chamber is in fluid communication with the separation chamber via a plurality of radially extending concentrate channels extending within the upper disk-shaped portion; the plurality of radially extending concentrate channels extend radially between respective concentrate channel inlets located radially outward of each and every one of the separate liquid inlets; When the bowl rotates, the upper disk-shaped portion and the outer wall rotate relative to the supply pipe, the separated liquid discharge pipe, the concentrated liquid discharge pipe, the separated liquid centripetal pump, and the concentrated liquid centripetal pump, respectively; the plurality of concentrate channels have channel portions intermediate their respective concentrate channel inlets and the concentrate centripetal pumping chamber, the cross-sectional area of ​​which, perpendicular to the direction of concentrate flow through the channel portions, increases continuously as the cross-sectional area asymptotically approaches the perpendicular axis; and the plurality of radially extending concentrate channels comprising a plurality of angularly spaced individual concentrate channels; the channel portions of the plurality of concentrate channels beginning at each concentrate channel inlet and extending radially inward and upward from said concentrate channel inlet; having a constant cross-sectional width perpendicular to the direction of concentrate flow and a vertical height that varies with radial distance from said axis; the upper disk-shaped portion having a plurality of angularly spaced vertical openings, each vertical opening extending between a radial periphery of the separation chamber and a respective channel inlet; An apparatus characterized in that

4. 1. An apparatus having a structure releasably received within a rotatable centrifuge bowl, the structure disposed within the bowl operatively separating cells in a cell culture into a cell concentrate and a cell separation solution within an interior region of the structure, the apparatus comprising: In the operating position, this structure an upper disk-shaped portion; The bottom and a cylindrical core vertically intermediate said upper disk-shaped portion and said lower portion; a separation chamber disposed radially outward from and surrounding the core; an outer wall configured to operatively engage the bowl, extending in fluid-tight relation to said upper disk-shaped portion and bounding said separation chamber; extending in a surrounding relationship to the core and the separation chamber; an outer wall having a frusto-conical interior shape, the inner radius of the outer wall being smaller adjacent the lower portion than the inner radius of the outer wall adjacent the upper disk-shaped portion; a vertically extending supply pipe; a separated liquid discharge pipe extending vertically; a concentrated liquid discharge pipe extending vertically; and the upper disk-shaped portion and the outer wall are rotatable about a vertical axis while operatively engaging the bowl; the structure further includes a centrifugal pump axially aligned with the core, coaxially positioned about the supply line, and in fluid communication with the centrate discharge line; the separate liquid centripetal pump is located within a separate liquid centripetal pump chamber within the upper disk-shaped portion; the separator centripetal pumping chamber is in fluid communication with the separator chamber via at least one separator channel extending within the upper disk-shaped portion; each separation channel extends in fluid communication between a separation channel inlet located radially outward of the core and the separation centripetal pumping chamber; the structure further includes a concentrate centripetal pump axially aligned with the core, coaxially positioned about the supply line, positioned vertically above the centrate centripetal pump, and in fluid communication with the concentrate discharge line; the concentrate centripetal pump is located within a concentrate centripetal pump chamber within the upper disk-shaped portion; the concentrate centripetal pumping chamber is in fluid communication with the separation chamber via a plurality of radially extending concentrate channels extending within the upper disk-shaped portion; the plurality of radially extending concentrate channels extend radially between respective concentrate channel inlets located radially outward of each and every one of the separate liquid inlets; When the bowl rotates, the upper disk-shaped portion and the outer wall rotate relative to the supply pipe, the separated liquid discharge pipe, the concentrated liquid discharge pipe, the separated liquid centripetal pump, and the concentrated liquid centripetal pump, respectively; the plurality of concentrate channels have channel portions intermediate their respective concentrate channel inlets and the concentrate centripetal pumping chamber, the cross-sectional area of ​​which, perpendicular to the direction of concentrate flow through the channel portions, increases continuously as the cross-sectional area asymptotically approaches the perpendicular axis; and the plurality of radially extending concentrate channels comprising a plurality of angularly spaced individual concentrate channels; the channel portions of the plurality of concentrate channels beginning at each concentrate channel inlet and extending radially inward and upward from said concentrate channel inlet; having a constant cross-sectional width perpendicular to the direction of concentrate flow and a vertical height that varies with radial distance from said axis; The upper disk-shaped portion has mating upper and lower pieces, the plurality of concentrate channels abutting respective surfaces of the upper and lower pieces. An apparatus characterized in that

5. 5. The device of any one of claims 1 to 4, wherein the channel portions of the plurality of concentrate channels begin at the respective concentrate channel inlets and extend radially inward.

6. 5. The device of any one of claims 1 to 4, wherein the channel portions of the plurality of concentrate channels begin at the respective concentrate channel inlets and extend radially inward and upward from the concentrate channel inlets.

7. the channel portions of the plurality of concentrate channels originate at the respective concentrate channel inlets and extend radially inward and upward from the concentrate channel inlets; The device of any one of claims 1 to 4, wherein the plurality of concentrate channels further have horizontally and radially extending portions, the horizontally and radially extending portions extending to a fluid midpoint between the channel portions and the concentrate centripetal pump chamber.

8. the channel portions of the plurality of concentrate channels originate at the respective concentrate channel inlets and extend radially inward and upward from the concentrate channel inlets; the plurality of concentrate channels further have horizontally and radially extending portions, the horizontally and radially extending portions extending fluidly intermediate the channel portions and the concentrate centripetal pumping chamber; the horizontally radially extending portions of the plurality of concentrate channels terminate radially inward at respective cell concentrate channel outlets within the concentrate centripetal pumping chamber; 5. The device of claim 1, wherein the horizontally radially extending portions of the plurality of concentrate channels have a constant cross-sectional area along their entire length.

9. the channel portions of the plurality of concentrate channels originate at the respective concentrate channel inlets and extend radially inward and upward from the concentrate channel inlets; 5. The apparatus of claim 1, wherein the plurality of concentrate channels further have horizontally radially extending portions, the horizontally radially extending portions extending radially outward from the concentrate centripetal pumping chamber.

10. the channel portions of the plurality of concentrate channels originate at the respective concentrate channel inlets and extend radially inward and upward from the concentrate channel inlets; the plurality of concentrate channels further have horizontally and radially extending portions, the horizontally and radially extending portions extending outward from the concentrate centripetal pumping chamber; the channel portion terminates in the horizontally and radially extending portion; 5. The device of claim 1, wherein the horizontally radially extending portions of the plurality of concentrate channels have a constant cross-sectional area along their entire length.

11. the channel portions of the plurality of concentrate channels originate at the respective concentrate channel inlets and extend radially inward and upward from the concentrate channel inlets; the plurality of concentrate channels further have horizontally and radially extending portions extending outward from each cell concentrate channel outlet into the concentrate centripetal pumping chamber; the concentrate centripetal pump has a concentrate centripetal pump inlet, each cell concentrate channel outlet being axially and radially aligned with the concentrate centripetal pump inlet; said plurality of concentrate channels terminating radially inward at their respective horizontally radially extending portions; 5. The device of claim 1, wherein the horizontally radially extending portions of the plurality of concentrate channels have a constant cross-sectional area along their entire length.

12. the upper disk-shaped portion having an upper piece and a lower piece in mating relationship therewith; 5. The device of any one of claims 1 to 4, wherein said plurality of concentrate channels abuts at least one lower surface of said upper piece and at least one upper surface of said lower piece.

13. 5. The device of any one of claims 1 to 4, wherein said plurality of radially extending concentrate channels comprises a single, substantially annular concentrate channel.

14. the plurality of radially extending concentrate channels comprises a single, substantially annular concentrate channel; 5. The device of any one of claims 1 to 4, wherein the channel portion of the concentrate channel begins at a substantially annular concentrate channel inlet and extends radially inward and upward from the concentrate channel inlet.

15. the plurality of radially extending concentrate channels comprises a single, substantially annular concentrate channel; the channel portion of the concentrate channel begins at a substantially annular concentrate channel inlet and extends radially inward and upward from the concentrate channel inlet; the concentrate channel further has a substantially annular horizontally and radially extending portion extending outward from the substantially annular cell concentrate channel outlet to the concentrate centripetal pumping chamber; 5. A device according to any one of claims 1 to 4, wherein the channel portion terminates radially inwardly at said horizontally radially extending portion.

16. the plurality of radially extending concentrate channels comprises a single, substantially annular concentrate channel; a channel portion of the concentrate channel beginning at a substantially annular concentrate channel inlet and extending radially inward and upward from the concentrate channel inlet; 5. The device of claim 1, wherein the upper disk-shaped portion has a substantially annular funnel channel extending upwardly and radially inwardly to the annular concentrate channel inlet.

17. the plurality of radially extending concentrate channels comprises a single, substantially annular concentrate channel; the channel portion of the concentrate channel begins at a substantially annular concentrate channel inlet and extends radially inward and upward from the concentrate channel inlet; the upper disk-shaped portion having a substantially annular funnel channel extending upwardly and radially inwardly to the annular concentrate channel inlet; the upper disk-shaped portion has a substantially annular cell concentrate guide surface that extends below the annular funnel channel and abuts radially outwardly against the separation chamber; 5. The device of claim 1, wherein the annular cell concentrate guiding surface further extends radially outward and upwardly adjacent the annular funnel channel.

18. the plurality of radially extending concentrate channels comprises a single, substantially annular concentrate channel; a channel portion of the concentrate channel beginning at a substantially annular concentrate channel inlet and extending radially inward and upward from the concentrate channel inlet; the upper disk-shaped portion having a substantially annular funnel channel extending upwardly and radially inwardly to the annular concentrate channel inlet; the upper disk-shaped portion has a substantially annular cell concentrate guide surface that extends below the annular funnel channel and abuts radially outwardly against the separation chamber; the annular cell concentrate guide surface further extends radially outward and upwardly adjacent the annular funnel channel; 5. The device of claim 1, wherein the upper disk-shaped portion is bounded on the lower side of the separation chamber by a radially extending surface, the radially extending surface terminating radially outward in a substantially annular edge, the annular edge axially above the radially outwardly extending annular cell concentrate guiding surface, and the annular funnel channel extends upward from the annular edge.

19. The device according to any one of claims 1 to 4, wherein the structure is a disposable structure.

20. 1. An apparatus having a structure releasably received within a rotatable centrifuge bowl, the structure disposed within the bowl operatively separating cells in a cell culture into a cell concentrate and a cell separation solution within an interior region of the structure, the apparatus comprising: In the operating position, this structure an upper disk-shaped portion; a cylindrical core extending vertically below the upper disk-like portion; an outer wall configured to operatively engage the bowl, extending in fluid-tight engagement with said upper disc-shaped portion; extending in surrounding relationship to the core; the end of the structure spaced vertically from the upper disk-like portion has a truncated conical shape with a smaller inner radius; an outer wall extending in surrounding relationship to the core and bounding a separation chamber within the structure extending radially intermediate the core and the outer wall; vertically extending cell culture supply piping; a separated liquid discharge pipe extending vertically; a concentrated liquid discharge pipe extending vertically; and the upper disk-shaped portion and the outer wall are rotatable about a vertical axis while operatively engaged with the bowl, and the supply pipe, the centrate discharge pipe, and the concentrate discharge pipe are coaxial with respect to the vertical axis; The upper disk-shaped portion a separate liquid centripetal pumping chamber in fluid communication with the separation chamber through at least one separate liquid opening; a concentrate centripetal pumping chamber in fluid communication with the separation chamber via a plurality of concentrate channels, the plurality of concentrate channels comprising: a concentrate channel inlet located radially outward from the at least one centrate opening; extending radially and fluidly between said plurality of concentrate channel inlets and said concentrate centripetal pumping chamber; the structure further comprises a separate liquid centripetal pump located coaxially with the supply line within the separate liquid centripetal pump chamber and in fluid communication with the separate liquid discharge line; a concentrate centripetal pump located coaxially with the supply line within the concentrate centripetal pump chamber and vertically above the centrate centripetal pump and in fluid communication with the concentrate discharge line; When the bowl rotates, the upper disk-shaped portion and the outer wall rotate relative to the supply pipe, the separated liquid discharge pipe, the concentrated liquid discharge pipe, the separated liquid centripetal pump, and the concentrated liquid centripetal pump, respectively; the plurality of concentrate channels have channel portions intermediate their respective concentrate channel inlets and the concentrate centripetal pumping chamber, the cross-sectional area of ​​which, perpendicular to the direction of concentrate flow through the channel portions, increases continuously as the cross-sectional area asymptotically approaches the perpendicular axis; and the plurality of concentrate channel portions originating at a respective concentrate channel inlet of each concentrate channel and extending radially inward and upward from the respective channel inlet; the plurality of concentrate channels comprising a plurality of angularly spaced individual concentrate channels; An apparatus characterized in that

21. 1. An apparatus having a structure for releasably receiving within a rotatable centrifuge bowl, the structure being disposed within the bowl and operatively separating cells in a cell culture into a concentrate and a clarifier within an interior region of the structure, the apparatus comprising: In the operating position, this structure an upper disk-shaped portion; The bottom and a cylindrical core vertically intermediate said upper disk-shaped portion and said lower portion; a separation chamber disposed radially outward from and surrounding the core; an outer wall configured to operatively engage the bowl, extending in operative fluid-tight relation to said upper disk-shaped portion and bounding said separation chamber; extending in a surrounding relationship to the core and the separation chamber; an outer wall having a frusto-conical interior shape, the inner radius of the outer wall being smaller adjacent the lower portion than the inner radius of the outer wall adjacent the upper disk-shaped portion; a vertically extending supply pipe; a separated liquid discharge pipe extending vertically; a concentrated liquid discharge pipe extending vertically; and the upper disk-shaped portion and the outer wall are rotatable about a vertical axis while operatively engaging the bowl; The upper disk-shaped portion a separate liquid chamber axially aligned with the core, coaxially positioned about the supply line, and in fluid communication with a separate liquid discharge line; the separation chamber is in fluid communication with the separation chamber via at least one separation channel extending within the upper disc-shaped portion; each separation channel extends in fluid communication between a respective separation channel inlet located radially outward of said core and said separation chamber; The upper disk-shaped portion a concentrate chamber axially aligned with the core, coaxially positioned about the supply pipe, positioned vertically above the separator chamber, and in fluid communication with the concentrate discharge pipe; the concentrate chamber is in fluid communication with the separation chamber via a plurality of radially extending concentrate channels extending within the upper disk-shaped portion; the plurality of radially extending concentrate channels extend radially between respective concentrate channel inlets located radially outward of each and every one of the separate liquid inlets; When the bowl rotates, the upper disk-shaped portion and the outer wall rotate relative to the supply pipe, the separated liquid discharge pipe, and the concentrated liquid discharge pipe, respectively; the plurality of concentrate channels each having a channel portion intermediate the concentrate channel inlet and the concentrate chamber, the cross-sectional area of ​​which is perpendicular to the direction of concentrate flow through the channel portion increasing continuously as the cross-sectional area of ​​the channel portion increases asymptotically to the perpendicular axis; and the plurality of radially extending concentrate channels comprising a plurality of angularly spaced individual concentrate channels; An apparatus characterized in that

22. 1. An apparatus having a structure for releasably receiving within a rotatable centrifuge bowl, the structure being disposed within the bowl and operatively separating cells in a cell culture into a concentrate and a clarifier within an interior region of the structure, the apparatus comprising: In the operating position, this structure an upper disk-shaped portion; The bottom and a cylindrical core vertically intermediate said upper disk-shaped portion and said lower portion; a separation chamber disposed radially outward from and surrounding the core; an outer wall configured to operatively engage the bowl, extending in operative fluid-tight relation to said upper disk-shaped portion and bounding said separation chamber; extending in a surrounding relationship to the core and the separation chamber; an outer wall having a frusto-conical interior shape, the inner radius of the outer wall being smaller adjacent the lower portion than the inner radius of the outer wall adjacent the upper disk-shaped portion; a vertically extending supply pipe; a separated liquid discharge pipe extending vertically; a concentrated liquid discharge pipe extending vertically; and the upper disk-shaped portion and the outer wall are rotatable about a vertical axis while operatively engaging the bowl; The upper disk-shaped portion a separate liquid chamber axially aligned with the core, coaxially positioned about the supply line, and in fluid communication with a separate liquid discharge line; the separation chamber is in fluid communication with the separation chamber via at least one separation channel extending within the upper disc-shaped portion; each separation channel extends in fluid communication between a respective separation channel inlet located radially outward of said core and said separation chamber; The upper disk-shaped portion a concentrate chamber axially aligned with the core, coaxially positioned about the supply pipe, positioned vertically above the separator chamber, and in fluid communication with the concentrate discharge pipe; the concentrate chamber is in fluid communication with the separation chamber via a plurality of radially extending concentrate channels extending within the upper disk-shaped portion; the plurality of radially extending concentrate channels extend radially between respective concentrate channel inlets located radially outward of each and every one of the separate liquid inlets; When the bowl rotates, the upper disk-shaped portion and the outer wall rotate relative to the supply pipe, the separated liquid discharge pipe, and the concentrated liquid discharge pipe, respectively; the plurality of concentrate channels each having a channel portion intermediate the concentrate channel inlet and the concentrate chamber, the cross-sectional area of ​​which is perpendicular to the direction of concentrate flow through the channel portion increasing continuously as the cross-sectional area of ​​the channel portion increases asymptotically to the perpendicular axis; and the plurality of radially extending concentrate channels comprising a plurality of angularly spaced individual concentrate channels; the channel portions of the plurality of concentrate channels beginning at each concentrate channel inlet and extending radially inward and upward from said concentrate channel inlet; a constant cross-sectional width in a direction perpendicular to the direction of concentrate flow, and a vertical height that varies with radial distance from said axis; An apparatus comprising:

23. 1. An apparatus having a structure for releasably receiving within a rotatable centrifuge bowl, the structure being disposed within the bowl and operatively separating cells in a cell culture into a concentrate and a clarifier within an interior region of the structure, the apparatus comprising: In the operating position, this structure an upper disk-shaped portion; The bottom and a cylindrical core vertically intermediate said upper disk-shaped portion and said lower portion; a separation chamber disposed radially outward from and surrounding the core; an outer wall configured to operatively engage the bowl, extending in operative fluid-tight relation to said upper disk-shaped portion and bounding said separation chamber; extending in a surrounding relationship to the core and the separation chamber; an outer wall having a frusto-conical interior shape, the inner radius of the outer wall being smaller adjacent the lower portion than the inner radius of the outer wall adjacent the upper disk-shaped portion; a vertically extending supply pipe; a separated liquid discharge pipe extending vertically; a concentrated liquid discharge pipe extending vertically; and the upper disk-shaped portion and the outer wall are rotatable about a vertical axis while operatively engaging the bowl; The upper disk-shaped portion a separate liquid chamber axially aligned with the core, coaxially positioned about the supply line, and in fluid communication with a separate liquid discharge line; the separation chamber is in fluid communication with the separation chamber via at least one separation channel extending within the upper disc-shaped portion; each separation channel extends in fluid communication between a respective separation channel inlet located radially outward of said core and said separation chamber; The upper disk-shaped portion a concentrate chamber axially aligned with the core, coaxially positioned about the supply pipe, positioned vertically above the separator chamber, and in fluid communication with the concentrate discharge pipe; the concentrate chamber is in fluid communication with the separation chamber via a plurality of radially extending concentrate channels extending within the upper disk-shaped portion; the plurality of radially extending concentrate channels extend radially between respective concentrate channel inlets located radially outward of each and every one of the separate liquid inlets; When the bowl rotates, the upper disk-shaped portion and the outer wall rotate relative to the supply pipe, the separated liquid discharge pipe, and the concentrated liquid discharge pipe, respectively; the plurality of concentrate channels each having a channel portion intermediate the concentrate channel inlet and the concentrate chamber, the cross-sectional area of ​​which is perpendicular to the direction of concentrate flow through the channel portion increasing continuously as the cross-sectional area of ​​the channel portion increases asymptotically to the perpendicular axis; and the plurality of radially extending concentrate channels comprising a plurality of angularly spaced individual concentrate channels; the channel portions of the plurality of concentrate channels beginning at each concentrate channel inlet and extending radially inward and upward from said concentrate channel inlet; having a constant cross-sectional width perpendicular to the direction of concentrate flow and a vertical height that varies with radial distance from said axis; the upper disk-shaped portion having a plurality of angularly spaced vertical openings, each vertical opening extending between a radial periphery of the separation chamber and a respective channel inlet; An apparatus characterized in that

24. 1. An apparatus having a structure for releasably receiving within a rotatable centrifuge bowl, the structure being disposed within the bowl and operatively separating cells in a cell culture into a concentrate and a clarifier within an interior region of the structure, the apparatus comprising: In the operating position, this structure an upper disk-shaped portion; The bottom and a cylindrical core vertically intermediate said upper disk-shaped portion and said lower portion; a separation chamber disposed radially outward from and surrounding the core; an outer wall configured to operatively engage the bowl, extending in operative fluid-tight relation to said upper disk-shaped portion and bounding said separation chamber; extending in a surrounding relationship to the core and the separation chamber; an outer wall having a frusto-conical interior shape, the inner radius of the outer wall being smaller adjacent the lower portion than the inner radius of the outer wall adjacent the upper disk-shaped portion; a vertically extending supply pipe; a separated liquid discharge pipe extending vertically; a concentrated liquid discharge pipe extending vertically; and the upper disk-shaped portion and the outer wall are rotatable about a vertical axis while operatively engaging the bowl; The upper disk-shaped portion a separate liquid chamber axially aligned with the core, coaxially positioned about the supply line, and in fluid communication with a separate liquid discharge line; the separation chamber is in fluid communication with the separation chamber via at least one separation channel extending within the upper disc-shaped portion; each separation channel extends in fluid communication between a respective separation channel inlet located radially outward of said core and said separation chamber; The upper disk-shaped portion a concentrate chamber axially aligned with the core, coaxially positioned about the supply pipe, positioned vertically above the separator chamber, and in fluid communication with the concentrate discharge pipe; the concentrate chamber is in fluid communication with the separation chamber via a plurality of radially extending concentrate channels extending within the upper disk-shaped portion; the plurality of radially extending concentrate channels extend radially between respective concentrate channel inlets located radially outward of each and every one of the separate liquid inlets; When the bowl rotates, the upper disk-shaped portion and the outer wall rotate relative to the supply pipe, the separated liquid discharge pipe, and the concentrated liquid discharge pipe, respectively; the plurality of concentrate channels each having a channel portion intermediate the concentrate channel inlet and the concentrate chamber, the cross-sectional area of ​​which is perpendicular to the direction of concentrate flow through the channel portion increasing continuously as the cross-sectional area of ​​the channel portion increases asymptotically to the perpendicular axis; and the plurality of radially extending concentrate channels comprising a plurality of angularly spaced individual concentrate channels; the channel portions of the plurality of concentrate channels beginning at each concentrate channel inlet and extending radially inward and upward from said concentrate channel inlet; having a constant cross-sectional width perpendicular to the direction of concentrate flow and a vertical height that varies with radial distance from said axis; The upper disk-shaped portion has mating upper and lower pieces, the plurality of concentrate channels abutting respective surfaces of the upper and lower pieces. An apparatus characterized in that

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