Apparatus and method for clean transfer of centrifuged material

Abstract

A fluid transfer container includes a tube, the tube including a conical portion at one end, the tube defining an interior space, and one or more fluid passages configured to allow fluid flow into and out of the interior space of the tube. The transfer container also includes a connector configured to seal the end of the conical portion only when the connector is joined to the conical portion, and a pressure control mechanism configured to increase or decrease pressure within the interior space of the tube. The transfer container can be used in a method that includes connecting a fluid line to the fluid passage, adding a fluid including a cryoprotectant and cells, centrifuging the transfer container, and then connecting the transfer container to a second fluid transfer container including another fluid. This allows for sterile transfer of the fluid and separation of the cells from the cryoprotectant.

Description

[Technical Field]

[0001] This disclosure claims priority to U.S. Provisional Patent Application No. 63 / 237,416, filed August 26, 2021.

[0002] The present disclosure relates to an apparatus and method for the clean transfer of centrifuged material, particularly conical tubes and / or syringes, and its use in handling bioprocess fluids. [Background technology]

[0003] Bioprocess fluids may be frozen during some processing or storage steps. Bioprocess fluids may contain cryoprotectants that help preserve cells or other components of the bioprocess fluid during freezing. While cryoprotectants can preserve cells or other components during the freezing process, they may be toxic to cells or adversely affect components of interest in the fluid when the fluid is thawed. Currently, ordinary conical tubes and syringes can be used to separate the cryoprotectant from other components of the bioprocess fluid. However, current methods allow air to come into contact with the bioprocess fluid and cannot guarantee sterility by themselves. Summary of the Invention

[0004] The present disclosure relates to an apparatus and method for the clean transfer of centrifuged material, particularly conical tubes and / or syringes, and its use in handling bioprocess fluids.

[0005] By using tubing connections and connectors, e.g., swabbable valves, conical tubes and / or syringes according to embodiments can enable the transfer of cells or other components of interest from one medium (e.g., a medium containing a cryoprotectant) to another environment (e.g., another medium without a cryoprotectant), which can simplify the handling process, such as reducing the risk of contamination and avoiding the use of a glove box or other such sterile environment to perform fluid transfers after thawing.

[0006] In one embodiment, the fluid transfer system includes a first fluid transfer container. The first fluid transfer container includes a tube, the tube including a conical portion at one end, the tube defining an interior space. The first fluid transfer container also includes one or more fluid passages configured to allow fluid flow into and out of the interior space of the tube. The first fluid transfer container further includes a connector disposed at the end of the conical portion, the connector configured to seal the end of the conical portion only when the connector is mated to the conical portion. The first fluid transfer container further includes a pressure control mechanism configured to increase or decrease the pressure within the interior space of the tube.

[0007] In one embodiment, the connector includes at least a portion of a swabbable valve.

[0008] In one embodiment, the fluid transfer system further includes a cap configured to cover the connector and at least a portion of the conical portion, hi one embodiment, the cap is configured to be received in a centrifuge.

[0009] In one embodiment, the fluid transfer system further includes a transfer assembly and a fluid line connected to the transfer assembly and one of the one or more fluid passages.

[0010] In one embodiment, the fluid transfer system further includes a medium supply and a fluid line connected to the medium supply and one of the one or more fluid passages.

[0011] In one embodiment, the fluid transfer system further includes a vent including a filter, and a fluid line connected to the vent and one of the one or more fluid passages.

[0012] In one embodiment, the pressure control mechanism includes a plunger movably disposed within the tube. In this embodiment, the one or more fluid passages are disposed on the plunger, the plunger including a socket, and the pressure control mechanism further includes a plunger handle configured to attach to the plunger in the socket. The plunger handle is configured such that a space is formed between the plunger handle and the tube, the space being configured to accommodate one or more fluid lines configured to be joined to the one or more fluid passages.

[0013] In one embodiment, the pressure control mechanism is a syringe including a syringe tube, a plunger, and a luer connector, the luer connector configured to connect to a transfer container luer connector provided on the transfer container.

[0014] In one embodiment, the fluid transfer system further includes a second fluid transfer container. The second fluid transfer container includes a second tube having a second conical portion at one end, the second tube defining a second interior space, and a second connector disposed at the end of the second conical portion. The connector is configured to seal the end of the second conical portion only when mated to the second conical portion. The connector of the first fluid transfer container and the second connector are configured to be mated to each other.

[0015] In one embodiment, the connector of the first fluid transfer container forms a first portion of a swabbable valve and the second connector forms a second portion of the swabbable valve.

[0016] In one embodiment, a method for handling fluids includes connecting one or more fluid lines to a fluid passageway of a first fluid transfer container. The method further includes directing a first fluid through at least one of the one or more fluid lines to the transfer container, the first fluid comprising previously frozen cells and a first culture medium, the first culture medium comprising a cryoprotectant, and centrifuging the first fluid transfer container while the first fluid transfer container contains the first fluid. The method also includes introducing a second fluid into the second fluid transfer container, the second fluid comprising a second culture medium, the second culture medium comprising a lower concentration of cryoprotectant than the first culture medium. The method further includes connecting the first fluid transfer container to a second fluid transfer container and transferring at least a portion of the first fluid to the second fluid transfer container.

[0017] In one embodiment, the method further includes placing a cap on the first connector prior to centrifugation of the syringe.

[0018] In one embodiment, connecting the first fluid transfer container to the second fluid transfer container includes mating a first connector to a second connector of the second fluid transfer container, hi one embodiment, the first connector is a first portion of a swabbable valve and the second connector is a second portion of the swabbable valve.

[0019] In one embodiment, the second medium does not contain any cryoprotectants.

[0020] In one embodiment, the method further includes directing the previously frozen cells and the second culture medium from the second fluid transfer container by pressurizing the interior space of the second fluid transfer container, the interior space of the second fluid transfer container being pressurized by driving a plunger disposed within the interior space of the second fluid transfer container.

[0021] In one embodiment, the method further includes directing the previously frozen cells and second culture medium from the second fluid transfer container by pressurizing the interior space of the second fluid transfer container, the interior space of the second fluid transfer container being pressurized by actuating the plunger of a syringe, the syringe being fluidly connected to the second fluid transfer container by a Luer connection. [Brief explanation of the drawings]

[0022] [Figure 1] 1 illustrates an apparatus for the transfer of fluids according to one embodiment. [Figure 2] 1 shows an apparatus for the transfer of fluids according to another embodiment; [Figure 3] 1 illustrates a system for the transfer of fluids according to one embodiment. [Figure 4] 1 shows a flowchart of a method for clean transfer of a fluid according to one embodiment. [Figure 5A] 1 illustrates a system for transferring fluid during filling of a device for fluid transfer according to one embodiment. [Figure 5B] 1 illustrates an apparatus for the transfer of fluids according to one embodiment when containing a fluid prior to centrifugation. [Figure 5C] 1 illustrates an apparatus for the transfer of fluids according to one embodiment when containing fluids after centrifugation. [Figure 5D] 1 illustrates an apparatus for the transfer of a fluid when included in a system for the transfer of a fluid according to one embodiment. [Figure 5E] 5D shows a second device for fluid transfer included in the system of FIG. 5C after fluid transfer. DETAILED DESCRIPTION OF THE INVENTION

[0023] The present disclosure relates to an apparatus and method for the clean transfer of centrifuged material, particularly conical tubes and / or syringes, and its use in handling bioprocess fluids.

[0024] FIG. 1 illustrates an apparatus for transferring fluids according to one embodiment. The apparatus 100 includes a body 102 having a tapered tip 104 and a connector 106 at the end of the tip 104. The apparatus 100 also includes a first fluid passageway 108, a second fluid passageway 110, and a pressure control connector 112. A pressure source 114 can be connected to the pressure control connector 112. In the embodiment illustrated in FIG. 1, the first fluid passageway 108, the second fluid passageway 110, and the pressure control connector 112 are formed in an end cap 116 attached to the end of the body 102 opposite the tapered tip 104. A cover 118 can be provided on the apparatus 100.

[0025] Device 100 can be used as part of a fluid transfer system. In one embodiment, the fluid transfer system can include multiple identical devices, such as device 100. In one embodiment, the fluid transfer system can further include one or more of a fluid reservoir 120 and / or a vacuum source 122 connected to first fluid passage 108 or second fluid passage 110. In an embodiment, device 100 can be filled with thawing fluid for a bioreaction, used for centrifugation, and / or filled with replacement medium, and / or can receive a portion of a bioreaction fluid.

[0026] The body 102 defines an interior space capable of containing a fluid. The body 102 can be made of any suitable material, such as a polymeric material. In one embodiment, the material used for the body 102 can be selected based on compatibility with the fluid to be handled using the device 100. The body 102 can include at least a portion having a cylindrical shape. The body 102 includes a tapered tip 104 at one end. The tapered tip 104 is provided at the end of the body 104. In one embodiment, the tapered tip 104 has a conical shape.

[0027] The end of the tapered tip 104 is provided with a connector 106. The connector 106 can be configured to selectively allow fluid to be drawn into or forced out of the body 102. In one embodiment, the connector 106 can be configured to allow fluid to be drawn into or forced out of the body 102 only when the connector 106 is mated to a corresponding connector. In one embodiment, the connector 106 includes at least a portion of a swabbable valve. In one embodiment, the connector 106 can include a check valve or a sterile connector. In one embodiment, the connector 106 of a first device 100 can be connected to a corresponding connector of a second device (shown in FIG. 3 and described below) so that fluid can be exchanged between the devices.

[0028] First fluid passageway 108 is a fluid passageway configured to allow fluid to pass into and out of device 100. First fluid passageway 108 may include one or more tube retention features that allow tubing to be joined to first fluid passageway 108. The tube retention features may include one or more sets of hose barbs, luer connections, at least part of a quick disconnect system, or any other suitable feature for retaining tubing so that the tubing can supply or receive fluid from first fluid passageway 108.

[0029] The second fluid passageway 110 is another fluid passageway similarly configured to allow fluid to enter and exit the device 100. The second fluid passageway 110 can include one or more tube retention features. The tube retention features can include one or more sets of hose barbs, luer connections, at least part of a quick disconnect system, or any other suitable features for retaining tubing so that the tubing can supply fluid to or receive fluid from the second fluid passageway 110. In one embodiment, the second fluid passageway 110 can have the same structure as the first fluid passageway 108. In one embodiment, the second fluid passageway 110 can differ from the first fluid passageway 108 in one or more of the length, diameter, provided connection features, or any other suitable feature.

[0030] The pressure control connector 112 is a connector configured to form a connection with the pressure source 114 so that the pressure source 114 can increase or decrease the pressure within the body 102. The pressure source 114 is a source of positive or negative pressure that can be connected to the device 100 via the pressure control connector 112. In one embodiment, the pressure source 114 includes a syringe that can provide positive pressure when the plunger is depressed or negative pressure when the plunger is retracted. In one embodiment, the pressure source 114 can be a vacuum, a pump, or any other suitable source of positive and / or negative pressure. The pressure source 114 and the pressure control connector 112 can be used to draw fluid into or expel fluid from the body 102. In one embodiment, the pressure control connector 112 is a Luer connection. In one embodiment, the pressure control connector 112 includes a quick-disconnect feature that allows for quick attachment and detachment of the pressure source 114. In one embodiment, the pressure control connector is an opening that can accept the tip of a tube from the pressure source 114.

[0031] The end cap 116 can surround the end of the body 102 opposite the tapered tip 104. The end cap 116 can be joined to the body 102 by any suitable temporary or permanent connection, such as a weld, or a mechanical connection such as a threaded connection, a snap fit, or the like. The connection of the end cap 116 to the body 102 can be a sealed connection, such as a weld as described above, or by including a seal in the mechanical connection. In one embodiment, the end cap 116 can include the first fluid passageway 108, the second fluid passageway 110, and / or the pressure control connector 112.

[0032] A cap 118 may optionally be included with device 100. Cap 118 is configured to cover at least the connector when attached to device 100. In one embodiment, cap 118 may further cover at least a portion of tapered tip 104 when attached to device 100. Cap 118 may be configured to be received in a centrifuge such that device 100 can be securely held in the centrifuge and the contents of device 100 can be separated using the centrifuge. Cap 118 may be configured to fit securely to device 100, for example, forming an interference or snap fit with body 102, for example at tapered tip 104.

[0033] Optionally, the device 100 can be connected to a fluid reservoir 120 via one of the first fluid passage 108 or the second fluid passage 110. In one embodiment, the connection can be made by attaching tubing from the fluid reservoir 120 to a tubing holder, such as a hose barb, luer connection, or the like, in the first fluid passage 108 or the second fluid passage 110. The fluid reservoir 120 can be configured to store or receive a bioreaction fluid. In one embodiment, the fluid reservoir 120 is a bag containing a bioreaction fluid that has been thawed after cryogenic storage or a processing step. In one embodiment, the thawed bioreaction fluid can include a cryoprotectant, a culture medium, and cells. In one embodiment, the fluid reservoir 120 contains a culture medium for cells in the thawed bioreaction fluid that does not include a cryoprotectant. In one embodiment, the fluid reservoir 120 can include multiple fluid sources that are combined within the device 100.

[0034] Optionally, device 100 can be connected to a vacuum source 122 through one of first fluid passageway 108 or second fluid passageway 110. The connection can be made by attaching tubing from vacuum source 122 to a tubing holder provided in first fluid passageway 108 or second fluid passageway 110. In one embodiment, vacuum source 122 can include a filter. In one embodiment, vacuum source 122 can be used to provide a vacuum to draw fluid from fluid reservoir 120 into body 102. In one embodiment, vacuum source 122 can be replaced with a vent device that allows pressure to be released as fluid is transferred from fluid reservoir 120 into body 102.

[0035] 2 illustrates an apparatus for transferring fluids according to another embodiment. The apparatus 200 includes a body 202 including a tapered tip 204 and a connector 206. The apparatus 200 further includes a plunger assembly 208 including a plunger 210, one or more seals 212, a mechanical connection interface 214 on the plunger 210, a shaft 216, and a plunger handle 218. A first fluid passage 220 and a second fluid passage 222 can be provided in the plunger 210. A cap 224 can comprise the apparatus 200. The apparatus 200 can optionally be connected to one or more of a fluid reservoir 226 and / or a vacuum source 228.

[0036] Device 200 can be used as part of a fluid transfer system. In one embodiment, the fluid transfer system can include multiple identical devices, such as device 200, or other devices described herein, including, by way of non-limiting example, device 100 shown in FIG. 1 above. In one embodiment, the fluid transfer system can further include one or more of a fluid reservoir 226 and / or a vacuum source 228 connected to first fluid passage 220 or second fluid passage 222. In an embodiment, device 200 can be filled with thawing fluid for a bioreaction, used for centrifugation, and / or filled with replacement medium, and / or can receive a portion of a bioreaction fluid.

[0037] The body 202 at least partially defines an interior space capable of containing a fluid. The body 202 can be configured to accommodate the plunger 210 such that the plunger 210 is slidable within the body 202, and the plunger 210 and the body 202 together define an interior space capable of containing a fluid. The body 202 can be made of any suitable material, such as a polymeric material. In one embodiment, the material used for the body 202 can be selected based on compatibility with the fluid to be handled using the device 200. The body 202 can include at least a portion having a cylindrical shape. The body 202 includes a tapered tip 204 at one end. The tapered tip 204 is provided at the end of the body 104. In one embodiment, the tapered tip 204 has a conical shape.

[0038] The end of the tapered tip 204 is provided with a connector 206. The connector 206 can be configured to selectively allow fluid to be drawn into or forced out of the body 202. In one embodiment, the connector 206 can be configured to allow fluid to be drawn into or forced out of the body 202 only when the connector 206 is mated to a corresponding connector. In one embodiment, the connector 206 includes at least a portion of a swabbable valve. In one embodiment, the connector 206 can include a check valve or a sterile connector. In one embodiment, the connector 206 of the first device 200 can be connected to a corresponding connector of a second device (shown in FIG. 3 and described below) so that fluid can be exchanged between the devices.

[0039] The plunger assembly 208 is an assembly that allows the plunger 210 to be positioned within a space defined by the body 202 and the movement of the plunger 210 therein. The plunger assembly 208 includes the plunger 210, one or more seals 212, a shaft 216, and a plunger handle 218.

[0040] The plunger 210 is configured to fit within the body 202 such that the plunger 210 and the body 202 define an interior space capable of containing a fluid. The plunger 210 is slidable within the body 202 such that the volume of the interior space can be adjusted by movement of the plunger 210, for example, to draw fluid into or expel fluid from the interior space. The plunger 210 can include a first fluid passageway 220 and / or a second fluid passageway 222 passing therethrough to allow fluid to enter or exit the interior space defined by the plunger 210 and the body 202. The first fluid passageway 220 and / or the second fluid passageway 222 can include hose retentions configured to allow a fluid line to be joined to the respective fluid passageways 220, 222. The hose retentions can be any suitable connection for a fluid line. As a non-limiting example, the hose retentions can include a hose barb.

[0041] The plunger 210 includes a mechanical connection interface 214. The mechanical connection interface 214 is any suitable interface for joining the shaft 216 to the plunger 210 by a mechanical connection. The mechanical connection may be a connection that can be made when the plunger 210 is disposed within the body 202 and / or while fluid lines are connected to the first fluid passageway 220 and / or the second fluid passageway 222. By way of non-limiting example, the mechanical connection of the shaft 216 to the plunger 210 may be provided by a snap, a detent, a thread, or the like, with corresponding features for such connection being provided on respective portions of the shaft 216 and the mechanical connection interface 214 of the plunger 210. In one embodiment, the mechanical connection interface 214 is a threaded connector configured to receive and engage a threaded end of the shaft 216.

[0042] One or more seals 212 may be provided to form a seal between the body 202 and the plunger 210. The seal may be any suitable seal, such as an O-ring, a gasket, or the like. The seal 212 may be provided in a channel formed around the plunger 210 facing the body 202. In one embodiment, there is one seal 212 between the plunger 210 and the body 202. In one embodiment, there are multiple seals between the plunger body 210 and the body 202. When multiple seals 212 are used, the seals may be positioned to reduce rocking or twisting of the plunger 210 along the path the plunger 210 may slide through the body 202. In one embodiment, the seals 212 may be distributed along the surface of the plunger 210 in a direction corresponding to the longitudinal direction of the body 202.

[0043] The shaft 216 is a shaft for the plunger assembly 208. When coupled to the plunger 210, the shaft 216 allows for the transfer of force to the plunger 210 so that the plunger 210 can move within the body 202. The shaft 216 can be sized smaller than the interior space of the portion of the body 202 through which the plunger 210 can move. In one embodiment, the shaft 216 is sized such that fluid lines connected to the first fluid passageway 220 and the second fluid passageway 222 can extend from the body 202 without interfering with the shaft 216. The shaft 216 can be configured to be mechanically coupled to the plunger 210 at the mechanical connection interface 214, for example, by including suitable features, such as threads, detents, flanges, etc., on the end of the shaft 216 configured to be received in the mechanical connection interface 214.

[0044] A plunger handle 218 may be provided on the end of the shaft 216 opposite the plunger 210. The plunger handle 218 may provide an area with which the plunger assembly 208 may engage to move the plunger 210 within the body 202. In one embodiment, the plunger handle 218 may be integrally formed with the shaft 216. In one embodiment, the plunger handle 218 may be connected to the shaft 216 by any suitable method, such as any suitable mechanical connection, adhesive, welding, or the like. The plunger handle 218 may have a diameter larger than the diameter of the shaft 216.

[0045] A cap 224 may optionally be used with device 200. Cap 224 may be used to cover connector 206 and optionally part or all of tapered tip 204, for example, during shipping or centrifugation of device 200. Cap 224 may be configured to be received in a centrifuge so that device 200 can be securely held in the centrifuge and the contents of device 200 can be separated using the centrifuge. Cap 224 may be configured to fit securely to device 200, for example, forming an interference or snap fit with body 202, for example at tapered tip 204.

[0046] Optionally, device 200 can be connected to a fluid reservoir 226 via one of first fluid passageway 220 or second fluid passageway 222. In one embodiment, the connection can be made by attaching tubing from fluid reservoir 226 to a tubing holder, such as a hose barb, luer connection, or the like, in first fluid passageway 220 or second fluid passageway 222. Fluid reservoir 226 can be configured to store or receive a bioreaction fluid. In one embodiment, fluid reservoir 226 is a bag containing a bioreaction fluid that has been thawed after cryogenic storage or a processing step. In one embodiment, the thawed bioreaction fluid can include a cryoprotectant, a culture medium, and cells. In one embodiment, fluid reservoir 226 contains a culture medium for cells in the thawed bioreaction fluid that does not include a cryoprotectant. In one embodiment, fluid reservoir 226 can include multiple separate fluid sources that are combined within device 200.

[0047] Optionally, device 200 can be connected to a vacuum source 228 through one of first fluid passageway 220 or second fluid passageway 222. The connection can be made by attaching tubing from vacuum source 228 to a tubing holder in first fluid passageway 220 or second fluid passageway 222. In one embodiment, vacuum source 228 can include a filter. In one embodiment, vacuum source 228 can be used to provide a vacuum to draw fluid from fluid reservoir 226 into body 202. In one embodiment, vacuum source 228 can be replaced with a vent device that allows pressure to be released as fluid is transferred from fluid reservoir 226 into body 202.

[0048] FIG. 3 illustrates a system for transferring fluids according to one embodiment. The system 300 includes a first fluid transfer device 310 and a second fluid transfer device 350. In the embodiment illustrated in FIG. 3, the first fluid transfer device 310 and the second fluid transfer device 350 are each the device 100 illustrated in FIG. 1 above. The connector 312 of the first fluid transfer device 310 and the connector 352 of the second fluid transfer device 350 allow fluid to be transferred between the first fluid transfer device 310 and the second fluid transfer device 350. In one embodiment, the connectors 312, 352 form a swabbable valve that allows fluid flow through each of the connectors 312, 352 only when the connectors 312 are connected to each other. 3, the transfer of fluid between the first fluid transfer device 310 and the second fluid transfer device 350 can be driven by applying positive or negative pressure to the interior spaces of the first fluid transfer device 310 and / or the second fluid transfer device 350 by a first pressure source 314 connected to a pressure control connector 316 on the first fluid transfer device 310 and / or a second pressure source 354 connected to a pressure control connector 356 on the second fluid transfer device 350. In one embodiment, the devices 310 and 350 can instead be devices according to the device 200 shown in FIG. 2 described above, where the application of positive or negative pressure is achieved by operating the plunger assembly 208 to increase or decrease the volume of the interior space defined by the body 202 and the plunger 210.

[0049] 4 shows a flowchart of a method for clean transfer of a fluid according to one embodiment. Method 400 includes connecting 402 one or more fluid lines to a fluid passageway of a first fluid transfer container, directing 404 a first fluid to the transfer container via at least one of the one or more fluid lines, the first fluid comprising previously frozen cells and a first culture medium, the first culture medium comprising a cryoprotectant, centrifuging 406 the first fluid transfer container while it contains the first fluid, introducing 408 a second fluid into the second fluid transfer container, and connecting 410 the first fluid transfer container to the second fluid transfer container. A portion of the first fluid can then be directed 412 from the first fluid transfer container to the second fluid transfer container. Method 400 can optionally further include 414 pushing the fluid out of the second fluid transfer container.

[0050] Method 400 includes connecting 402 one or more fluid lines to a fluid passageway of a first fluid transfer container. The fluid passageway may be, for example, fluid passageway 108 or 110 shown in FIG. 1 and described above, or fluid passageway 220 or 222 shown in FIG. 2 and described above. The connection may be made by any suitable fluid line connection provided on the fluid transfer container, such as a hose barb, luer connection, etc. The connection allows fluid to be introduced into the interior space of the first fluid transfer container via the one or more fluid lines.

[0051] The method 400 further includes, at 404, directing a first fluid into the transfer container via at least one of the one or more fluid lines. The first fluid includes previously frozen cells and a first culture medium. The first culture medium includes a cryoprotectant. The cryoprotectant can be any suitable cryoprotectant compound used in freezing cells. The fluid can be provided from one or more fluid sources, such as a bag containing previously frozen cells and culture medium, a reservoir containing additional culture medium, etc.

[0052] The first fluid transfer vessel, when it contains the first fluid, is centrifuged 406. The first fluid transfer vessel can be inserted into a centrifuge and spun to separate the contents of the fluid contained within the first fluid transfer vessel. Optionally, a cap can be placed on the first fluid transfer vessel. The cap can protect features of the first fluid transfer vessel, such as the connector, and can facilitate inserting and retaining the first fluid transfer vessel in the centrifuge, etc. Centrifugation can separate the first fluid into a cell-rich fraction containing a relatively high percentage of thawed cells and a medium-rich fraction containing a relatively high percentage of the first medium. The cell-rich fraction can be formed in a portion of the first fluid transfer vessel proximate a connector, such as connector 106 or 206, as shown above in FIGS. 1 and 2, respectively.

[0053] The method 400 can further include introducing 408 a second fluid into the second fluid transfer container. The second fluid includes a second culture medium. The second culture medium can be any suitable medium for the previously frozen cells contained in the first fluid. The second culture medium includes a lower concentration of cryoprotectant than the first culture medium in the first fluid transfer container. In one embodiment, the second culture medium does not include any cryoprotectant.

[0054] At 410, the first fluid transfer container is connected to the second fluid transfer container. The connection can be made via a connector provided on each of the fluid transfer containers, such as connectors 106 and 206 shown above in Figures 1 and 2, respectively. The connectors can be configured to allow flow only when the two connectors are connected to one another. In one embodiment, the connectors of the first and second fluid transfer containers form a swabbable valve that allows fluid to pass between the first and second fluid transfer containers when the connectors of the first and second fluid transfer containers are connected.

[0055] A portion of the first fluid can then be directed from the first fluid transfer container to a second fluid transfer container at 412. The first fluid passes through the connection formed at 410 to pass from the first fluid transfer container to the second fluid transfer container. The portion of the first fluid directed from the first fluid transfer container to the second fluid transfer container can include some or all of the cell-rich fraction obtained by centrifugation at 406. Optionally, the portion of the first fluid transferred from the first fluid transfer container to the second fluid transfer container includes a portion of the medium-rich fraction in addition to the cell-rich fraction. The portion of the first fluid transferred from the first fluid transfer container to the second fluid transfer container can be mixed with the second medium in the second fluid transfer container. This results in mixing of the thawed cells and a portion of the first medium with the second medium, and therefore the thawed cells are in a medium having a lower concentration of cryoprotectant compared to when the cells were in the first fluid.

[0056] Optionally, the fluid in the second fluid transfer vessel can be removed at 414. The fluid can be removed from the second fluid transfer vessel to continue a process, such as a biological reaction process. The fluid can be removed from the second fluid transfer vessel by, for example, connecting a fluid line to fluid passages 108 or 110 shown in FIG. 1 and described above, or fluid passages 220 or 222 shown in FIG. 2 and described above, and using those fluid connections to remove the fluid from the second transfer vessel. The removed fluid can include culture medium and thawed cells.

[0057] 5A shows a system for transferring fluids during filling of a device for fluid transfer according to one embodiment. A first fluid transfer container 500 is connected to a container of thawed cells 502 and a medium source 504 by a first fluid passage 506. A vacuum source 508 is connected to a second fluid passage 508. This allows both medium from medium source 504 and thawed cells from 502 to be introduced into the interior space of first fluid transfer container 500.

[0058] The first fluid transfer container 500 can then be disconnected from the thawed cell container 502, the medium source 504, and the vacuum source 508. Figure 5B shows an apparatus for fluid transfer according to one embodiment when it contains fluid prior to centrifugation. The medium from the medium source 504 and the thawed cells from 502 shown in Figure 1 are present together in a first mixture 512.

[0059] The first fluid transfer container 500 can then be centrifuged. Figure 5C shows an apparatus for fluid transfer according to one embodiment when it contains fluid after centrifugation. Centrifugation separates the first mixture 512 into a cell-rich fraction 514 and a medium-rich fraction 516, with the cell-rich fraction 514 containing a relatively high number of thawed cells and the medium-rich fraction 516 containing a relatively high number of medium.

[0060] 5D shows an apparatus for fluid transfer as it would appear in a system for fluid transfer according to one embodiment. A first fluid transfer container 500 containing a cell-rich fraction 514 and a medium-rich fraction 516 is connected to a second fluid transfer container 520 via a connector 518 on the first fluid transfer container 500 and a connector 522 on a second fluid transfer container 520. The connectors 518 and 522 can be mated to allow fluid to flow from the first fluid transfer container 500 to the second fluid transfer container 520, for example, by opening a swabbable valve when the connectors 518, 522 are mated. At least a portion of the contents of the first fluid transfer container 500 can be drained from the first fluid transfer container 500 to the second fluid transfer container 520. The portion drained from the first fluid transfer container can include some or all of the cell-rich fraction 514 and, optionally, a portion of the medium-rich fraction 516. Ejection of the portion can be achieved by attaching a pressure source, such as a syringe, pressurized fluid line, or the like, to the pressure control connector 524 of the first fluid transfer container 500 and applying positive pressure to the interior space of the first fluid transfer container 500. In one embodiment, the first fluid transfer container can instead be a device such as device 200 shown in FIG. 2, and ejection of the portion can be achieved by depressing plunger 210 to reduce the volume of the interior space defined by plunger 210 and body 202, as shown in FIG. 2 above. In one embodiment, ejection of the portion from the first fluid transfer container 500 can be achieved by, for example, retracting the plunger to increase the volume of the interior space, by providing negative pressure to the second fluid transfer container 520 to draw fluid into the second fluid transfer container, or by providing negative pressure via a pressure source connected to the pressure control connector 526 of the second fluid transfer container 520. The fluid ejected from the first fluid transfer container 500 can be mixed with the low-temperature cryoprotectant medium 528 contained within the second fluid transfer container 520. Low temperature cryoprotectant medium 528 may be a medium suitable for thawing cells from 502 that does not contain a cryoprotectant or that contains a cryoprotectant at a concentration lower than the concentration of the cryoprotectant that was present in first mixture 512.

[0061] 5E shows the second fluid transfer container 520 after transfer of fluid from the first fluid transfer container 500. The first fluid transfer container 500 and any remaining fluid contained therein, e.g., medium-rich fraction 516, may be discarded, or optionally, the first fluid transfer container 500 may be flushed and cleaned for subsequent use. The second fluid transfer container 520 contains cells 530 from the cell-rich portion 514 dispersed in low-temperature cryoprotectant medium 528.

[0062] Aspects: It is understood that any of the embodiments 1-11 can be combined with any of the embodiments 12-18.

[0063] Aspect 1. a first fluid transfer vessel, a tube including a conical portion at one end and defining an interior space; one or more fluid passages configured to allow fluid flow into or out of the interior space of the tube; a first fluid transfer vessel comprising: a connector disposed at an end of the conical portion and configured to seal the end of the conical portion only when joined to the conical portion; a pressure control mechanism configured to increase or decrease the pressure in the interior space of the tube; A fluid transfer system comprising:

[0064] Embodiment 2. The fluid transfer system of embodiment 1, wherein the connector comprises at least a portion of a swabbable valve.

[0065] Embodiment 3. The fluid transfer system of embodiment 1 or 2, further comprising a cap configured to cover the connector and at least a portion of the conical portion.

[0066] Embodiment 4. The fluid transfer system of embodiment 3, wherein the cap is configured to be received in a centrifuge.

[0067] Embodiment 5. The fluid transfer system of any of embodiments 1 to 4, further comprising a transfer assembly and a fluid line connected to the transfer assembly and one of the one or more fluid passages.

[0068] Embodiment 6. A fluid transfer system according to any of embodiments 1 to 5, further comprising a medium supply and a fluid line connected to the medium supply and to one of the one or more fluid passages.

[0069] Embodiment 7. The fluid transfer system of any of embodiments 1 to 6, further comprising a vent including a filter, and a fluid line connected to the vent and one of the one or more fluid passages.

[0070] Aspect 8: The pressure control mechanism includes a plunger movably disposed within the tube; one or more fluid passages disposed in the plunger; the plunger includes a socket; the pressure control mechanism further includes a plunger handle configured to be attached to the plunger at the socket, the plunger handle configured such that a space is formed between the plunger handle and the tube, the space configured to accommodate one or more fluid lines configured to be joined to the one or more fluid passages; 8. The fluid transfer system of any one of embodiments 1 to 7.

[0071] Aspect 9. A fluid transfer system described in any of aspects 1 to 8, wherein the pressure control mechanism is a syringe including a syringe tube, a plunger, and a luer connector, and the luer connector is configured to be connected to a transfer container luer connector provided on the transfer container.

[0072] Aspect 10: a second tube including a second conical portion at one end and defining a second interior space; a second connector disposed at an end of the second conical portion and configured to seal the end of the second conical portion only when joined to the second conical portion; and a second fluid transfer vessel comprising: The connector of the first fluid transfer container and the second connector are configured to be joined to each other; 10. The fluid transfer system of any one of embodiments 1 to 9.

[0073] Aspect 11: A fluid transfer system as described in aspect 11, wherein the connector of the first fluid transfer container forms a first portion of the swabbable valve and the second connector forms a second portion of the swabbable valve.

[0074] Embodiment 12. A method of handling a fluid, comprising: connecting one or more fluid lines to a fluid passageway of a first fluid transfer vessel; directing a first fluid to a transfer container via at least one of the one or more fluid lines, the first fluid comprising previously frozen cells and a first culture medium, the first culture medium comprising a cryoprotectant; Centrifuging the first fluid transfer vessel when the first fluid transfer vessel contains the first fluid; introducing a second fluid into a second fluid transfer vessel, the second fluid comprising a second culture medium, the second culture medium comprising a lower concentration of cryoprotectant than the first culture medium; connecting the first fluid transfer vessel to a second fluid transfer vessel; transferring at least a portion of the first fluid to a second fluid transfer vessel; A method comprising:

[0075] Embodiment 13. The method of embodiment 12, further comprising placing a cap on the first connector prior to centrifugation of the syringe.

[0076] Embodiment 14. The method of embodiment 12 or 13, wherein connecting the first fluid transfer container to the second fluid transfer container comprises mating the first connector to a second connector of the second fluid transfer container.

[0077] Aspect 15: The method of aspect 14, wherein the first connector is a first portion of a swabbable valve and the second connector is a second portion of the swabbable valve.

[0078] Embodiment 16: The method according to any of embodiments 12 to 15, wherein the second culture medium does not comprise any cryoprotectant.

[0079] Embodiment 17. The method of any of embodiments 12 to 16, further comprising directing the previously frozen cells and the second culture medium from the second fluid transfer vessel by pressurizing an interior space of the second fluid transfer vessel, wherein the interior space of the second fluid transfer vessel is pressurized by driving a plunger disposed within the interior space of the second fluid transfer vessel.

[0080] Embodiment 18. The method of any of embodiments 12 to 17, further comprising directing the previously frozen cells and the second culture medium from the second fluid transfer container by pressurizing an interior space of the second fluid transfer container, wherein the interior space of the second fluid transfer container is pressurized by driving a plunger of a syringe, and the syringe is fluidly connected to the second fluid transfer container by a Luer connection.

[0081] The examples disclosed in this application should be considered in all respects as illustrative and not restrictive. The scope of the invention is indicated by the appended claims, rather than the foregoing description, and all changes that come within the meaning and range of equivalency of the claims are intended to be embraced therein.

Claims

1. 1. A fluid transfer system comprising a first fluid transfer vessel, the first fluid transfer vessel comprising: a tube including a conical portion at one end and defining an interior space; one or more fluid passages configured to allow fluid flow into or out of the interior space of the tube; a first connector disposed at an end of the conical portion and configured to seal the end of the conical portion only when joined to the conical portion; a pressure control mechanism configured to increase or decrease the pressure in the interior space of the tube; Including, the first fluid transfer container is configured to be centrifuged; A fluid transfer system, wherein the first connector is configured to be joined to the second connector of the second fluid transfer container to allow fluid to flow from the first fluid transfer container to the second fluid transfer container after the first fluid transfer container has been centrifuged.

2. A fluid transfer system as described in claim 1, wherein the first connector includes at least a portion of a swabbable valve.

3. A fluid transfer system as described in claim 1, further comprising a cap configured to cover at least a portion of the conical portion and the first connector.

4. The fluid transfer system of claim 1 , further comprising a vent including a filter, and a fluid line connected to the vent and one of the one or more fluid passages.

5. 1. A method of handling a fluid, comprising: connecting one or more fluid lines to one or more fluid passages of a first fluid transfer vessel; directing a first fluid through at least one of the one or more fluid lines to a first fluid transfer vessel, the first fluid comprising previously frozen cells and a first culture medium, the first culture medium comprising a cryoprotectant; Centrifuging the first fluid transfer vessel when the first fluid transfer vessel contains the first fluid; introducing a second fluid into a second fluid transfer vessel, the second fluid comprising a second culture medium, the second culture medium comprising a lower concentration of cryoprotectant than the first culture medium; connecting the first fluid transfer vessel to a second fluid transfer vessel after centrifuging the first fluid transfer vessel; transferring at least a portion of the first fluid to a second fluid transfer vessel; Including, a first fluid transfer vessel comprising: a tube including a conical portion at one end and defining an interior space; one or more fluid passages configured to allow fluid flow into or out of the interior space of the tube; a first connector disposed at an end of the conical portion and configured to seal the end of the conical portion only when joined to the conical portion; a pressure control mechanism configured to increase or decrease the pressure in the interior space of the tube; Including, a second fluid transfer container including a second connector; A method wherein connecting a first fluid transfer vessel to a second fluid transfer vessel comprises mating a first connector and a second connector to one another.

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