A piston-type biological liquid separation cup with a core and a separation method thereof

Abstract

The present invention provides a piston-type biological liquid separation cup with a core, the separation cup comprising a liquid inlet, a core, a separation chamber and a liquid outlet; the liquid inlet and the liquid outlet are separately arranged; the biological liquid enters the separation chamber through the liquid inlet and the core; after the biological liquid is separated in the separation chamber, part of the liquid is discharged through the liquid outlet, and the remaining liquid is discharged through the core and the liquid inlet. The present invention also provides a separation method for a piston-type biological liquid separation cup with a core. The piston-type biological liquid separation cup with a core and the separation method provided by the present invention can continuously process biological liquids with high efficiency, and the amount of liquid processed at a single time is not limited; the centrifugal path is long, the centrifugal time is sufficient, the centrifugal effect is better, and the cell recovery rate is higher; the inlet and outlet are separated, and the liquid inlet, separation chamber, diversion chamber, liquid outlet, etc. can be cleaned, thereby avoiding the loss of target cells.

Description

Technical Field

[0001] The invention belongs to the field of biological liquid separation, and in particular relates to a piston-type biological liquid separation cup with a core and a separation method thereof. Background Art

[0002] In modern biotechnology, it is often necessary to process biological fluids to obtain one or several target cells. For example, remove excess plasma and red blood cells from peripheral blood and umbilical cord blood to obtain stem cells or immune cells; or concentrate and wash the cultured cell fluid to remove excess waste fluid and obtain cell fluid of target concentration. Since the volume of biological fluids is generally not fixed, such as the volume of umbilical cord blood to be processed is generally between 40ml-150ml, and the volume of cell fluid to be concentrated and washed is generally between 200ml-50L, for the above-mentioned biological fluids with non-fixed volumes, the current popular method is to use a "piston-type" separation cup with a variable volume. For example, patent number CN1331610A discloses a system for separating components of biological fluids, in which the matching "piston-type" separation cup is described in detail. The separation cup structure includes a hollow centrifugal treatment chamber, which has an axial biological fluid inlet / outlet; the treatment chamber has a movable piston, which "sucks" a fixed amount of biological fluid into the treatment chamber under the action of the "air pump" of its supporting equipment. After the treatment is completed, the piston moves upward and squeezes out the treated biological fluid components through the outlet; the piston position is monitored by the optical device of the supporting equipment, and the pressure regulating valve device on the equipment is used to selectively connect the treatment chamber and the container, or cut off their connection. Patent No. CN109294899A discloses "a method for preparing PBMC cells", which also includes a "piston centrifugal cylinder" that cooperates with the pneumatic device on the equipment to suck / exhaust biological fluid. Its structure is basically the same as the structure of the separation cup disclosed in Patent No. CN1331610A.

[0003] The piston-type separation cup can flexibly process biological fluids with variable volumes, but the separation cups of current technology have the following problems:

[0004] (1) The amount of liquid processed at a time is small, and biological liquids cannot be processed continuously. Since the amount of liquid processed at a time by the "piston-type" separation cup cannot exceed its maximum volume (generally 220ml-250ml), when the amount of biological liquid processed exceeds its maximum volume, multiple cycles must be performed; for example, if its maximum volume is 250ml, when the amount of liquid processed is 40ml-150ml of umbilical cord blood, only one cycle is required to complete the processing (one cycle includes aspiration of liquid → centrifugation → removal of waste liquid → collection of target cells); and when the amount of liquid processed is 2000ml of cell fluid, at least 8 cycles are required, which is very inefficient;

[0005] (2) The target cells near the inlet are wasted, reducing the target cell recovery rate. Since the inlet and outlet of the separation cup are the same, each time the biological fluid to be separated is inhaled, some will inevitably remain at the inlet. After the centrifugation is completed, the "waste liquid" will be discharged first, and the biological fluid containing target cells at the inlet will also be discharged together, wasting precious target cells. The more cycles, the more target cells will be wasted.

[0006] (3) The later the biological fluid is sucked in each cycle, the closer it is to the rotation axis, and the shorter the centrifugation time (relative to the beginning of sucking the biological fluid), and the less sufficient the centrifugation. Whether the biological fluid is centrifuged sufficiently is closely related to the centrifugation time and the centrifugal force; since the centrifugal force G = ω 2 r (where ω is the speed and r is the centrifugal radius). At the same speed, the centrifugal radius of the part of the liquid closer to the top axis of the separation cup is shorter, the centrifugal force is smaller, and the centrifugation is less complete. At the end of the centrifugation, a few target cells that have not been fully centrifuged in time will be mixed in the waste liquid and discharged together, resulting in a low recovery rate of target cells. Summary of the invention

[0007] In order to solve at least one of the above technical problems, the technical solution adopted by the present invention is to provide a piston-type biological liquid separation cup with a core and a separation method thereof, which can efficiently and continuously process biological liquids and the amount of liquid processed at a single time is not limited; the loss of target cells can be avoided; the centrifugation path is long, the centrifugation time is sufficient, the centrifugation effect is good, and the cell recovery rate is higher.

[0008] In order to achieve at least one of the above purposes, the technical solution adopted by the present invention is:

[0009] The present invention provides a piston-type biological liquid separation cup with a core, the separation cup comprising a liquid inlet, a core, a separation chamber and a liquid outlet; the liquid inlet and the liquid outlet are separately arranged; the biological liquid enters the separation chamber through the liquid inlet and the core; after the biological liquid is separated in the separation chamber, part of the liquid is discharged through the liquid outlet, and the rest of the liquid is discharged through the core and the liquid inlet.

[0010] Furthermore, the inner core is a hollow cylindrical rotating body, and the inner core includes a hemispherical focusing hole, a guide cone, an annular baffle, a waist-shaped hole and a strip hole; the biological liquid enters the separation chamber through the hemispherical focusing hole, the guide cone, the annular baffle and the waist-shaped hole in sequence, and the hemispherical focusing hole, the guide cone, the annular baffle and the waist-shaped hole constitute a guide channel; the strip hole connects the separation chamber and the liquid outlet fluid.

[0011] Furthermore, it also includes a cup body and a piston; the cup body is a hollow transparent cylinder, the core is fixed in the cup body, the piston is slidably arranged between the core and the cup body, and is slidingly sealed with the core and the cup body; the cup body, the core and the piston are coaxially arranged, and the cavity between the inner surface of the cup body, the drainage cone surface above the piston and the outer surface of the core forms the separation chamber.

[0012] Furthermore, a first flange and a second flange are provided on the outer side surface of the bottom of the cup body, and a stepped end surface is provided at the end surface of the bottom; it also includes a base, the base is disc-shaped, and an air inlet is provided at the bottom of the base, and a limiting ring, a limiting groove and a third flange are provided on the upper part of the base, and a plurality of small holes for ventilation are evenly distributed on the limiting ring, and the small holes connect the air inlet and the air pressure chamber fluid under the piston; a hollow annular bottom cover is provided on the outer side of the base, and a plurality of evenly distributed buckles are provided on the edge of the bottom cover, and the buckles cooperate with the second flange to fix the base to the bottom of the cup body.

[0013] Furthermore, it also includes a fixed head, which is a hollow multi-step columnar body, and the fixed head includes a first stepped shaft, a second stepped shaft and a circular disk; the liquid inlet is through-set on the first stepped shaft, and the liquid outlet is through-set on the second stepped shaft; the bottom of the circular disk is provided with a first convex ring, a second convex ring and a third convex ring, and an outer flange, a first groove and a second groove are sequentially provided between the first convex ring, the second convex ring and the third convex ring from the outside to the inside.

[0014] Furthermore, it also includes a first liner and a second liner, both of which are inverted funnel-shaped; the first liner includes a first small-end hollow cylinder and a first large-end cone, and the outer side of the first small-end hollow cylinder is provided with a step shaft and an annular groove; the inner surface of the first small-end hollow cylinder is axially provided with a plurality of first support bars evenly distributed; the second liner includes a second small-end hollow cylinder and a second large-end cone; the interior of the second small-end hollow cylinder is provided with a hollow step hole; the outer side surface of the second large-end cone is provided with a plurality of second support bars evenly distributed; a second dynamic sealing ring is provided on the annular groove of the first liner; the step shaft shape of the first small-end hollow cylinder on the first liner is adaptively inserted into the second stepped shaft of the fixed head and is abutted and fixed with the third convex ring; after the second small end hollow cylinder of the second liner passes through the first small end hollow cylinder, the stepped shaft at the end is bonded to the stepped hole of the fixed head, and the outer surface of the second small end hollow cylinder abuts the first support bar inside the first liner hollow cylinder, and at the same time, the second support bar of the second liner abuts the inner side of the first large end cone of the first liner; it also includes a guide tube, which is bonded to the hollow step hole; the guide tube, the inner surface of the first small end hollow cylinder and the liquid inlet form a liquid inlet channel; the strip hole, the outer side surface of the second large end cone, the inner surface of the first small end hollow cylinder, the outer side of the second small end hollow cylinder, the inner side of the first small end hollow cylinder and the liquid outlet constitute a liquid outlet channel.

[0015] Furthermore, it also includes a bearing seat, which is a cylindrical hollow rotating body, the upper part of the bearing seat includes an upper convex ring, a third groove, a through hole is arranged in the middle, and an upper step ring surface is arranged between the upper convex ring and the through hole; the lower part of the bearing seat is provided with an annular step surface; a first static sealing ring is arranged in the third groove, and a first dynamic sealing ring is arranged on the upper step ring surface; the upper convex ring is bonded to the first groove, and the first convex ring is bonded to the third groove; the first convex ring and the second convex ring are respectively abutted against the first static sealing ring and the first dynamic sealing ring.

[0016] Furthermore, it also includes a cup cover, which includes a third small-end hollow cylinder and a third large-end cone, and the bottom end face of the third large-end cone is provided with a fourth convex ring and a fourth groove; the upper end of the cup body is provided with a fifth convex ring and a fifth groove; the fourth convex ring is bonded in the fifth groove, and the fifth convex ring is bonded in the fourth groove.

[0017] Furthermore, it also includes a bearing, which is installed in the bearing hole of the bearing seat through an interference fit of the outer ring; the third small end hollow cylinder of the cup cover is interference fit and positioned with the inner ring of the bearing, and inserted into the second groove; and the outer surface of the third small end hollow cylinder has an interference fit with the first dynamic sealing ring, and a rotational seal is formed between the two; the inner surface of the third small end hollow cylinder has an interference fit with the second dynamic sealing ring, and a rotational seal is formed between the two.

[0018] The present invention also provides a separation method of the piston-type biological liquid separation cup with a core according to the above-mentioned claim, wherein the separation method includes single processing and continuous processing;

[0019] The steps of the single treatment are:

[0020] a. The centrifuge on the separation equipment drives the separation cup to rotate at high speed;

[0021] b. Using a peristaltic pump as the power, a certain amount of gradient liquid ficoll is transported to the hemispherical flow-collecting hole at the bottom of the inner core through the liquid inlet channel; under the action of centrifugal force, the gradient liquid quickly climbs along the diversion channel to the annular baffle, and then quickly climbs along the waist-shaped hole and the drainage cone surface to the separation chamber;

[0022] c Use a peristaltic pump to transport the biological fluid to be separated from the liquid inlet channel to the hemispherical flow-collecting hole at the bottom of the inner core; the biological fluid will quickly enter the separation chamber along the diversion channel; when the biological fluid completely enters the separation cup, all the biological fluid will be located in the separation chamber; after a period of centrifugation, the heavier red blood cells are located at the outermost side of the separation chamber, followed by the gradient fluid, the buffy coat layer and the plasma in turn;

[0023] d. Use an air pump to push the piston to gradually reduce the volume of the separation chamber, and push out the plasma and buffy coat layer inside in turn; cooperate with the valve, color sensor, pipeline component sensor, component collection bag and waste liquid bag on the separation equipment to collect the required components;

[0024] eWhen only red blood cells and gradient fluid are left in the separation chamber, the air pump reverses the pumping process to drive the piston downward, while the centrifuge slowly stops. The liquid in the separation chamber flows into the hemispherical flow-collecting hole under the action of gravity. The remaining liquid is sucked away from the flow guide tube under the action of the peristaltic pump, completing the separation.

[0025] The steps of the continuous treatment are:

[0026] A The centrifuge on the separation equipment drives the separation cup to rotate at high speed;

[0027] Device B uses a peristaltic pump as a power source to transport the biological fluid from the liquid inlet channel to the hemispherical flow-collecting hole at the bottom of the inner core; under the action of centrifugal force, the biological fluid quickly climbs along the diversion channel to the annular baffle, and then quickly climbs along the waist-shaped hole and the drainage cone surface to the separation chamber, and the cell fluid gradually fills the separation chamber; in the process of the biological fluid filling the separation chamber, the cells in the biological fluid are distributed on the outermost side of the separation chamber under the action of centrifugal force, while the waste liquid is at the innermost side of the separation chamber;

[0028] C With the continuous input of biological fluid, the waste liquid without cells on the innermost side of the separation chamber is squeezed out; when the biological fluid is completely input into the separation cup, the outer side of the separation chamber is cells and the inner side is waste liquid, and the total volume is the maximum dynamic volume of the separation cup;

[0029] D. Start the air pump and push the piston, cooperating with the gravity sensor, optical sensor, valve and other components on the separation equipment until a certain volume remains in the separation cup, and then stop pushing the piston;

[0030] E The centrifuge is stopped and the air pump is started to pump air in the reverse direction to move the piston to the bottom. Under the action of gravity, all the biological fluid in the separation cup is collected at the hemispherical flow-collecting hole at the bottom of the inner core. Then the peristaltic pump is started to suck away the remaining biological fluid from the guide tube.

[0031] F sucks a small amount of cleaning liquid from the liquid outlet and starts the centrifuge to clean the liquid outlet, liquid outlet channel, separation chamber and diversion channel; then starts the peristaltic pump again to suck the residual cells from the liquid inlet channel into the product bag.

[0032] Compared with the prior art, the piston-type biological liquid separation cup with a core and the separation method thereof provided by the present invention have the following beneficial effects:

[0033] (1) Compared with the common piston-type separation cup, by adding a core, a fluid channel from the inlet to the outlet is established in the separation cup, which can efficiently and continuously process biological fluids, and the amount of liquid processed at a time is not limited; it can be used to separate small volumes of biological fluids such as umbilical cord blood, peripheral blood, and bone marrow, and can also be used to concentrate and wash large volumes of cell fluids;

[0034] (2) By increasing the inner core, the minimum centrifugal force of the biological fluid in the separation chamber is limited, a better centrifugal effect is obtained, and the recovery rate of target cells will be higher;

[0035] (3) Establishing a fluid channel for the biological liquid to flow from the inlet of the separation cup, the central guide tube, the guide cavity to the bottom of the separation cavity, and then diffuse upward to the liquid outlet; from the inlet to the outlet, the centrifugal path is long, the centrifugal time is sufficient, the centrifugal effect is better, and the cell recovery rate is higher;

[0036] (4) The separation cup inlet and outlet are separated, and the liquid inlet, separation chamber, diversion chamber, liquid outlet, etc. can be cleaned, thereby avoiding the loss of target cells and improving the cell recovery rate. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 It is a structural schematic diagram of a piston-type biological liquid separation cup with a core according to the present invention;

[0038] Figure 2 It is a schematic diagram of an explosion of a piston-type biological liquid separation cup with a core according to the present invention;

[0039] Figure 3 It is a structural schematic diagram of the fixing head of the present invention;

[0040] Figure 4 It is a structural schematic diagram of the first liner of the present invention;

[0041] Figure 5 for Figure 4 Schematic diagram of the cross section along the middle line BB;

[0042] Figure 6 is a schematic structural diagram of the second liner of the present invention;

[0043] Figure 7 for Figure 6 The view from the middle M direction;

[0044] Figure 8 It is a structural schematic diagram of the bearing seat of the present invention;

[0045] Fig. 9 It is a structural schematic diagram of the cup cover of the present invention;

[0046] Fig.10 It is a schematic diagram of the structure of the cup body of the present invention;

[0047] Fig.11 It is a structural schematic diagram of the piston of the present invention;

[0048] Fig.12 It is a structural schematic diagram of the kernel of the present invention;

[0049] Fig.13 It is a structural schematic diagram of the base of the present invention;

[0050] Fig.14 It is a schematic diagram of the internal structure of the base of the present invention;

[0051] Figure 15-19 It is a schematic diagram of the process of the separation cup of the present invention when the volume of the separated biological fluid can be processed in a single time;

[0052] Figure 20-23It is a schematic diagram of the process of the separation cup of the present invention when the volume of separated biological fluid needs to be processed continuously.

[0053] Among them, 1 fixed head, 1A first stepped shaft, 1A1 stepped hole, 1B second stepped shaft, 1C circular disk, 1C1 first convex ring, 1C2 second convex ring, 1C3 third convex ring, 1C4 outer flange, 1C5 first groove, 1C6 second groove, 1D liquid inlet, liquid outlet 1E, 2 first static seal ring, 3 first dynamic seal ring, 4 bearing seat, 4A upper convex ring, 4B third groove, 4C through hole, 4D upper step ring surface, 4E annular step surface, 5 bearing, 6 cup cover, 6A third small end hollow cylinder, 6B third large end cone, 6B1 fourth convex ring, 6B2 fourth groove, 7 second dynamic seal ring, 8 first liner, 8A first small end hollow cylinder, 8A1 stepped shaft, 8A2 annular groove, 8B first large end cone, 8C first support bar, 9 second liner, 9A second small end hollow cylinder, 9A1 hollow step hole, 9B The second large end cone, 9B1 the second support bar, 10 the guide tube, 11 the inner core, 11A the first step column, 11B the second step column, 11C the third step column, 11D the step end surface, 11E the strip hole, 11F the waist hole, 11G the annular baffle, 11H the guide cone surface, 11J the hemispherical focusing hole, 12 the cup body, 12A the fifth convex ring, 12B the fifth groove, 12C the first flange, 12D the second flange, 1 2E stepped end face, 13 third piston sealing ring, 14 first piston sealing ring, 15 piston, 15A sixth groove, 15B seventh groove, 15C eighth groove, 15D ninth groove, 15E drainage cone, 15F air pressure chamber, 16 second piston sealing ring, 17 second static sealing ring, 18 base, 18A air inlet hole, 18B limiting ring, 18C limiting groove, 18D third flange, 18E small hole, 19 bottom cover. DETAILED DESCRIPTION

[0054] In order to enable those skilled in the art to better understand the technical solution of the present invention, the present invention is further described in detail below in conjunction with specific embodiments. Please note that the embodiments described below are exemplary and are only used to explain the present invention, and cannot be understood as limiting the present invention. If no specific technology or conditions are specified in the embodiments, the technology or conditions described in the literature in the art or the product instructions shall be followed.

[0055] In the description of the present invention, it should be understood that the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of the features. In the description of the present invention, the meaning of "plurality" is two or more, unless otherwise clearly and specifically defined.

[0056] In the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", "fixed" and the like should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0057] The following will describe in detail the piston-type biological liquid separation cup with a core and the separation method thereof provided by the present invention through specific embodiments:

[0058] As attached Figure 1-14 As shown, the piston-type biological liquid separation cup with a core provided by the present invention mainly includes a fixed head 1, a first static seal ring 2, a first dynamic seal ring 3, a bearing seat 4, a bearing 5, a cup cover 6, a second dynamic seal ring 7, a first liner 8, a second liner 9, a guide tube 10, a core 11, a cup body 12, a third piston seal ring 13, a first piston seal ring 14, a piston 15, a second piston seal ring 16, a second static seal ring 17, a base 18 and a bottom cover 19 and other components.

[0059] like Figure 3 As shown, the main body of the fixed head 1 is a hollow multi-step columnar body, including a first stepped shaft 1A, a second stepped shaft 1B, and a circular disc 1C. A hollow cylindrical liquid inlet 1D is provided through the first stepped shaft 1A, and a stepped hole 1A1 is also provided in the first stepped shaft 1A; a hollow cylindrical liquid outlet 1E is provided through the second stepped shaft 1B; a first convex ring 1C1, a second convex ring 1C2, and a third convex ring 1C3 are provided at the bottom of the circular disc 1C, and an outer flange 1C4, a first groove 1C5, and a second groove 1C6 are provided between the above-mentioned convex rings from the outside to the inside.

[0060] like Figure 4-5 As shown, the first liner 8 is in the shape of an inverted funnel, including a first small end hollow cylinder 8A and a first large end cone 8B. The outside of the first small end hollow cylinder 8A is provided with a step shaft 8A1 and an annular groove 8A2; the inner surface of the first small end hollow cylinder 8A is provided with three evenly distributed first support bars 8C along the axial direction.

[0061] like Figure 6-7 As shown, the second liner 9 is also in an inverted funnel shape, including a second small end hollow cylinder 9A and a second large end cone 9B. The second small end hollow cylinder 9A is provided with a hollow step hole 9A1 inside; and the outer surface of the second large end cone 9B is provided with three evenly distributed second support bars 9B1.

[0062] like Figure 8As shown, the bearing seat 4 is a cylindrical hollow rotating body. The upper part of the bearing seat 4 includes an upper convex ring 4A, a third groove 4B, a through hole 4C is arranged in the middle, and an upper step ring surface 4D is arranged between the upper convex ring 4A and the through hole 4C; the lower part of the bearing seat 4 is provided with an annular step surface 4E.

[0063] The annular groove 8A2 of the first liner 8 is provided with a second dynamic seal ring 7; the stepped shaft 8A1 of the first small-end hollow cylinder 8A on the first liner 8 is inserted into the second stepped shaft 1B of the fixed head 1 in a shape-fitting manner, and is fixedly abutted with the third convex ring 1C3. After the second small-end hollow cylinder 9A of the second liner 9 passes through the first small-end hollow cylinder 8A, the stepped shaft at the end is bonded to the stepped hole 1A1 of the fixed head 1, and the outer surface of the second small-end hollow cylinder 9A abuts against the three first support bars 8C inside the first liner hollow cylinder 8A, and at the same time, the three second support bars 9B1 of the second liner 9 abut against the inner side of the first large-end cone 8B of the first liner 8. The guide tube 10 is a hollow long round tube, and the outer wall and the upper end surface of the guide tube 10 are bonded to the hollow stepped hole 9A1 in the second liner 9. The bearing 5 is installed in the bearing hole of the bearing seat 4 through an interference fit of the bearing outer ring; the first static seal ring 2 is arranged in the third groove 4B of the bearing seat 4, and the first dynamic seal ring 3 is arranged on the upper step ring surface 4D of the bearing seat 4. The upper convex ring 4A of the bearing seat 4 is bonded to the first groove 1C5 of the fixed head 1, and the first convex ring 1C1 of the fixed head 1 is bonded to the third groove 4B of the bearing seat 4. The first convex ring 1C1 and the second convex ring 1C2 of the fixed head 1 are respectively pressed and contacted with the first static seal ring 2 and the first dynamic seal ring 3 installed on the bearing seat 4 to seal the fixed head 1 and the bearing seat 4. In summary, the fixed head 1, the first liner 8, the second liner 9, the guide tube 10 and the bearing seat 4 are fixedly connected together to form a stationary component of the separation cup, that is, the separation cup is in a stationary state when separating.

[0064] like Fig. 9 As shown, the cup cover 6 is an inverted funnel-shaped rotating body, including a third small end hollow cylinder 6A and a third large end cone 6B, and a fourth convex ring 6B1 and a fourth groove 6B2 are arranged on the bottom end surface of the third large end cone 6B.

[0065] like Fig.10 As shown, the cup body 12 is a hollow transparent cylinder. The upper end of the cup body 12 is provided with a fifth convex ring 12A and a fifth groove 12B matching the cup cover, and the inner side of the upper end is provided with a stepped cylindrical surface. The outer side surface of the bottom of the cup body 12 is provided with a first flange 12C and a second flange 12D, and the end surface of the bottom is provided with a stepped end surface 12E.

[0066] like Fig.11As shown, the piston 15 is a hollow cylindrical rotating body. The outer side of the piston 15 is provided with a sixth groove 15A and a seventh groove 15B for installing a sealing ring; the inner side of the piston 15 is provided with an eighth groove 15C and a ninth groove 15D for installing a sealing ring; the upper side of the piston 15 is provided with a conical surface 15E for drainage; and the bottom of the piston 15 is provided with a gas pressure chamber 15F.

[0067] like Fig.12 As shown, the inner core 11 is a stepped cylindrical shell. The inner core 11 comprises a first stepped column 11A, a second stepped column 11B and a third stepped column 11C; a stepped end surface 11D is provided between the first stepped column 11A and the second stepped column 11B, four uniformly distributed strip holes 11E are provided at the connection between the second stepped column 11B and the third stepped column 11C, uniformly distributed waist-shaped holes 11F are provided on the third stepped column 11C, and a circular baffle 11G, a guide cone 11H and a hemispherical focusing hole 11J are provided on the inner side of the third stepped column 11C.

[0068] like Figure 13-14 As shown, the base 18 is disc-shaped. An air inlet 18A is arranged at the bottom of the base 18, a limiting ring 18B, a limiting groove 18C and a third flange 18D are arranged at the top of the base 18, and four small holes 18E for ventilation are evenly distributed on the limiting ring 18B.

[0069] The first step column 11A, the second step column 11B and the step end face 11D of the inner core 11 are bonded to the step cylindrical surface of the cup body 12 and positioned. The first piston seal ring 14 and the second piston seal ring 16 are respectively installed in the sixth groove 15A and the seventh groove 15B of the piston 15; the third seal ring 13 is respectively installed in the eighth groove 15C and the ninth groove 15D of the piston 15. The piston 15 is slidably arranged on the inner core 11 and can slide between the inner core 11 and the cup body 12, and the first piston seal ring 14 and the second piston seal ring 16 are in contact with and sealed on the inner surface of the cup body 12, and the third seal ring 13 is in contact with and sealed on the outer surface of the inner core 11. The base 18 is arranged at the bottom of the cup body 12, wherein a second static sealing ring 17 is arranged on the stepped end face 12E at the bottom of the cup body 12, and the third flange 18D of the base 18 contacts the end face of the cup body 12 and presses the second static sealing ring 17; the bottom end face of the inner core 11 is inserted into the limiting groove 18C of the base 18. A hollow annular bottom cover 19 is arranged on the outside of the base 18, and a plurality of evenly distributed buckles are arranged on the edge of the bottom cover 19, which cooperate with the second flange 12D of the cup body 12 to fix the base 18 at the bottom of the cup body 12. The cup cover 6 is bonded to the upper part of the cup body 12, wherein the fourth convex ring 6B1 of the cup cover 6 is bonded to the fifth groove 12B of the cup body 12, and the fifth convex ring 12A of the cup body 12 is bonded to the fourth groove 6B2 of the cup cover 6. In summary, the cup cover 6, the inner core 11, the cup body 12, the piston 15, the base 18 and the bottom cover 19 constitute a rotating assembly of the separation cup, that is, the separation cup is in a high-speed rotating state when performing separation.

[0070] The third small end hollow cylinder 6A of the cup cover 6 in the rotating component is interference fit and positioned with the inner ring of the bearing 5, and is inserted into the second groove 1C6 of the fixed head 1; and the outer surface of the third small end hollow cylinder 6A has an interference fit with the first dynamic sealing ring 3, and the two can be rotatably sealed; the inner surface of the third small end hollow cylinder 6A has an interference fit with the second dynamic sealing ring 7, and the two can also be rotatably sealed; thereby realizing a rotating support and sealing connection between the rotating component and the stationary component through the bearing.

[0071] In the separation cup provided by the present invention, the guide tube 10, the inner surface of the hollow cylinder 8A at the first small end of the second liner 8 and the liquid inlet 1D at the top of the fixed head 1 form a liquid inlet channel; the hemispherical focusing hole 11J, the guide cone 11H, the annular baffle 11G and the waist-shaped hole 11F of the inner core 11 constitute the guide channel; the inner surface of the cup body 12, the guide cone 15E and the outer surface of the inner core 11 form a separation chamber. When the piston 15 is at the bottom, the dynamic volume of the separation cup is the largest, preferably 250ml; when the piston 15 moves upward, the separation chamber will gradually become smaller, and when the piston 15 moves upward, the separation chamber will gradually become smaller. When the plug 15 moves to the top, the dynamic volume of the separation cup is the smallest, preferably 30 ml; the strip hole 11E above the inner core 11, the outer side surface of the second large end cone 9B, the inner surface of the first small end hollow cylinder 8A, the outer side of the second small end hollow cylinder 9A and the inner side of the first small end hollow cylinder 8A and the liquid outlet 1E of the fixed head 1 constitute a liquid outlet channel; the air inlet hole 18A of the base 18, the small hole 18E on the limit ring 18B, and the air pressure chamber 15F at the bottom of the piston 15 together constitute the air inlet and outlet channels, and the air inlet and outlet channels are not connected to the separation chamber at any time. In summary, when the separation cup provided by the present invention is used for separation, the biological fluid is separated through the liquid inlet channel, the guide channel, the separation chamber and the liquid outlet channel in sequence, so that a fluid channel from the liquid inlet to the liquid outlet is established in the separation cup, the centrifugal path is long, the centrifugal time is sufficient, and the centrifugal effect is better; and the inlet and outlet separation setting can clean the liquid inlet, the separation chamber, the guide chamber, the liquid outlet, etc., thereby avoiding the loss of target cells; at the same time, by actuating the piston through the air inlet and outlet channels, the volume of the separation chamber can be changed, and the biological liquid can be continuously processed with high efficiency, and the amount of liquid processed at a single time is not limited.

[0072] The first static seal ring 2 and the second static seal ring 17 provided by the present invention are preferably ordinary rubber rings, preferably made of NBR; the first dynamic seal ring 3 and the second dynamic seal ring 7 are preferably fluororubber rings with smooth surfaces; the first piston seal ring 14, the second piston seal ring 16 and the third piston seal ring 13 are preferably silicone rings with smooth surfaces.

[0073] The separation method of the piston-type biological liquid separation cup with a core provided by the present invention is specifically described as follows:

[0074] 1. As attached Figure 15-19 As shown, when the volume of the biological fluid to be separated is between 40-250 ml, taking the separation of 120 ml of umbilical cord blood + 30 ml of gradient fluid as an example, the following steps are included:

[0075] a. The centrifuge on the separation equipment drives the rotating assembly of the separation cup to rotate at high speed;

[0076] b. Using a peristaltic pump or other power source as the power, a certain amount of gradient liquid ficoll is transported to the hemispherical flow-collecting hole at the bottom of the inner core through the liquid inlet channel; under the action of centrifugal force, the gradient liquid quickly climbs along the diversion channel to the annular baffle, and then quickly climbs along the waist-shaped hole and the drainage cone surface to the separation chamber;

[0077] c Use a peristaltic pump to transport the cord blood to be separated from the liquid inlet channel to the hemispherical flow collection hole at the bottom of the inner core; similarly, the cord blood will quickly enter the separation chamber along the diversion channel; when the cord blood completely enters the separation cup, all the cord blood will be located in the separation chamber. After a period of centrifugation, the heavier red blood cells are located at the outermost side of the separation chamber, followed by the gradient liquid, the buffy coat layer, and the plasma in turn;

[0078] d. Use an air pump to push the piston through the air inlet and outlet channels to gradually reduce the volume of the separation chamber, and push out the plasma and buffy coat layer inside in turn; cooperate with the valve, color sensor, pipeline component sensor, component collection bag and waste liquid bag on the separation equipment to collect the required components;

[0079] eWhen only red blood cells and gradient fluid are left in the separation chamber, the air pump reverses the pumping process to drive the piston downward, while the centrifuge slowly stops. The liquid in the separation chamber flows into the hemispherical flow-collecting hole under the action of gravity. Under the action of the peristaltic pump, the remaining liquid is sucked away from the flow guide tube to complete the separation.

[0080] 2. As attached Figure 20-23 As shown in the figure, when the volume of the biological fluid to be separated is greater than 250 ml, the cell concentration of 1000 ml is counted as 1×10 7 The cell solution was concentrated to a volume of 100 ml and the cell count was 1×10 8 Take pcs / ml as an example, including the following steps:

[0081] The centrifuge on the A separation device drives the rotating assembly of the separation cup to rotate at high speed;

[0082] Device B uses a peristaltic pump or other power source as a power source to transport the cell fluid from the liquid inlet channel to the hemispherical flow-collecting hole at the bottom of the inner core; under the action of centrifugal force, the cell fluid quickly climbs along the diversion channel to the annular baffle, and then quickly climbs along the waist-shaped hole and the drainage cone surface to the separation chamber, and the cell fluid gradually fills the separation chamber; in the process of the cell fluid filling the separation chamber, the cells in the cell fluid are distributed on the outermost side of the separation chamber under the action of centrifugal force, while the waste liquid is at the innermost side of the separation chamber;

[0083] C With the continuous input of cell fluid, the waste fluid without cells in the innermost part of the separation chamber is squeezed out; when the cell fluid is completely input into the separation cup, the outer side of the separation chamber is cells and the inner side is waste fluid, and the total volume is the maximum dynamic volume of the separation cup, 250ml;

[0084] D. Start the air pump, push the piston, and cooperate with the gravity sensor, optical sensor, valve and other components on the separation equipment until the remaining volume in the separation cup is 100 ml, then stop pushing the piston;

[0085] E. The centrifuge stops and the air pump is started to pump air in the reverse direction, causing the piston to move to the bottom. Under the action of gravity, all the cell fluid in the separation cup is collected at the hemispherical flow-collecting hole at the bottom of the inner core. Then the peristaltic pump is started to suck away the remaining cell fluid from the guide tube.

[0086] F sucks a small amount of cleaning liquid from the liquid outlet and starts the centrifuge to clean the liquid outlet, liquid outlet channel, separation chamber and diversion channel; then starts the peristaltic pump again to suck the residual cells from the liquid inlet channel into the product bag; completes the continuous concentration of biological liquid.

[0087] In the present invention, unless otherwise clearly specified and limited, a first feature being "above" or "below" a second feature may mean that the first and second features are in direct contact, or the first and second features are in indirect contact through an intermediate medium. Moreover, a first feature being "above", "above" or "above" a second feature may mean that the first feature is directly above or obliquely above the second feature, or simply means that the first feature is higher in level than the second feature. A first feature being "below", "below" or "below" a second feature may mean that the first feature is directly below or obliquely below the second feature, or simply means that the first feature is lower in level than the second feature.

[0088] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, unless they are contradictory.

Claims

1. A piston-type biological liquid separation cup with a core, characterized in that: The separation cup comprises a liquid inlet, a core, a separation chamber and a liquid outlet; the liquid inlet is separately arranged from the liquid outlet; the biological liquid enters the separation chamber through the liquid inlet and the core; After the biological liquid is separated in the separation chamber, part of the liquid is discharged through the liquid outlet, and the rest of the liquid is discharged through the inner core and the liquid inlet; The separation cup further comprises a cup body and a piston; the cup body is a hollow transparent cylinder, the core is fixed in the cup body, the piston is slidably disposed between the core and the cup body, and is in sliding sealing cooperation with the core and the cup body; the cup body, the core and the piston are coaxially arranged, and the cavity between the inner surface of the cup body, the drainage cone surface above the piston and the outer surface of the core forms the separation chamber; The inner core is a stepped cylindrical shell.

2. The piston-type biological liquid separation cup with a core according to claim 1, characterized in that: The inner core comprises a hemispherical focusing hole, a flow guiding cone, an annular baffle, a waist-shaped hole and a strip hole; the biological liquid enters the separation chamber through the hemispherical focusing hole, the flow guiding cone, the annular baffle and the waist-shaped hole in sequence, and the hemispherical focusing hole, the flow guiding cone, the annular baffle and the waist-shaped hole constitute a flow guiding channel; the strip hole connects the separation chamber and the liquid outlet fluid.

3. The piston-type biological liquid separation cup with a core according to claim 1, characterized in that: The outer side surface of the bottom of the cup body is provided with a first flange and a second flange, and the end surface of the bottom is provided with a stepped end surface; it also includes a base, the base is disc-shaped, the bottom of the base is provided with an air inlet hole, the upper part of the base is provided with a limiting ring, a limiting groove and a third flange, the limiting ring is evenly provided with a plurality of small holes for ventilation, the small holes connect the air inlet hole and the air pressure chamber fluid under the piston; a hollow annular bottom cover is provided on the outer side of the base, and a plurality of evenly distributed buckles are provided on the edge of the bottom cover, and the buckles cooperate with the second flange to fix the base to the bottom of the cup body.

4. The piston-type biological liquid separation cup with a core according to claim 2, characterized in that: It also includes a fixed head, which is a hollow multi-step columnar body, and includes a first stepped shaft, a second stepped shaft and a circular disk; the liquid inlet is through-set on the first stepped shaft, and the liquid outlet is through-set on the second stepped shaft; the bottom of the circular disk is provided with a first convex ring, a second convex ring and a third convex ring, and an outer flange, a first groove and a second groove are sequentially provided between the first convex ring, the second convex ring and the third convex ring from the outside to the inside.

5. The piston-type biological liquid separation cup with a core according to claim 4, characterized in that: It also includes a first liner and a second liner, both of which are inverted funnel-shaped; the first liner includes a first small-end hollow cylinder and a first large-end cone, and the outer side of the first small-end hollow cylinder is provided with a step shaft and an annular groove; the inner surface of the first small-end hollow cylinder is axially provided with a plurality of first support bars evenly distributed; the second liner includes a second small-end hollow cylinder and a second large-end cone; the interior of the second small-end hollow cylinder is provided with a hollow step hole; the outer side surface of the second large-end cone is provided with a plurality of second support bars evenly distributed; a second dynamic sealing ring is provided on the annular groove of the first liner; the step shaft shape of the first small-end hollow cylinder on the first liner is adapted to be inserted into the second stepped shaft of the fixed head, and is connected with The third convex ring is abutted and fixed; after the second small end hollow cylinder of the second liner passes through the first small end hollow cylinder, the stepped shaft at the end is bonded to the stepped hole of the fixed head, and the outer surface of the second small end hollow cylinder abuts the first support bar inside the first small end hollow cylinder, and at the same time, the second support bar of the second liner abuts the inner side of the first large end cone of the first liner; it also includes a guide tube, which is bonded to the hollow step hole; the guide tube, the inner surface of the first small end hollow cylinder and the liquid inlet form a liquid inlet channel; the strip hole, the outer side surface of the second large end cone, the inner surface of the first small end hollow cylinder, the outer side of the second small end hollow cylinder, the inner side of the first small end hollow cylinder and the liquid outlet constitute a liquid outlet channel.

6. The piston-type biological liquid separation cup with a core according to claim 5, characterized in that: It also includes a bearing seat, which is a cylindrical hollow rotating body. The upper part of the bearing seat includes an upper convex ring, a third groove, a through hole in the middle, and an upper step ring surface between the upper convex ring and the through hole; the lower part of the bearing seat is provided with an annular step surface; a first static sealing ring is provided in the third groove, and a first dynamic sealing ring is provided on the upper step ring surface; the upper convex ring is bonded to the first groove, and the first convex ring is bonded to the third groove; the first convex ring and the second convex ring are respectively abutted against the first static sealing ring and the first dynamic sealing ring.

7. The piston-type biological liquid separation cup with a core according to claim 6, characterized in that: It also includes a cup cover, which includes a third small-end hollow cylinder and a third large-end cone, and a fourth convex ring and a fourth groove are arranged on the bottom end face of the third large-end cone; a fifth convex ring and a fifth groove are arranged on the upper end of the cup body; the fourth convex ring is bonded in the fifth groove, and the fifth convex ring is bonded in the fourth groove.

8. The piston-type biological liquid separation cup with a core according to claim 7, characterized in that: It also includes a bearing, which is installed in the bearing hole of the bearing seat through an interference fit of the outer ring; the third small end hollow cylinder of the cup cover is interference fit and positioned with the inner ring of the bearing, and inserted into the second groove; and the outer surface of the third small end hollow cylinder has an interference fit with the first dynamic sealing ring, and a rotational seal is formed between the two; the inner surface of the third small end hollow cylinder has an interference fit with the second dynamic sealing ring, and a rotational seal is formed between the two.

9. A separation method of the piston-type biological liquid separation cup with a core according to any one of claims 1 to 8, wherein the separation method comprises single treatment and continuous treatment; The steps of the single treatment are: a. The centrifuge on the separation equipment drives the separation cup to rotate at high speed; b. Using a peristaltic pump as the power, a certain amount of gradient liquid ficoll is transported to the hemispherical flow-collecting hole at the bottom of the inner core through the liquid inlet channel; under the action of centrifugal force, the gradient liquid quickly climbs along the diversion channel to the annular baffle, and then quickly climbs along the waist-shaped hole and the drainage cone surface to the separation chamber; c Use a peristaltic pump to transport the biological fluid to be separated from the liquid inlet channel to the hemispherical flow-collecting hole at the bottom of the inner core; the biological fluid will quickly enter the separation chamber along the diversion channel; when the biological fluid completely enters the separation cup, all the biological fluid will be located in the separation chamber; after a period of centrifugation, the heavier red blood cells are located at the outermost side of the separation chamber, followed by the gradient fluid, the buffy coat layer and the plasma in turn; d. Use an air pump to push the piston to gradually reduce the volume of the separation chamber, and push out the plasma and buffy coat layer inside in turn; Cooperate with valves, color sensors, pipeline component sensors, component collection bags and waste liquid bags on the separation equipment to collect the required components; eWhen only red blood cells and gradient fluid are left in the separation chamber, the air pump reverses the pumping process to drive the piston downward, while the centrifuge slowly stops. The liquid in the separation chamber flows into the hemispherical flow-collecting hole under the action of gravity. The remaining liquid is sucked away from the flow guide tube under the action of the peristaltic pump, completing the separation. The steps of the continuous treatment are: A The centrifuge on the separation equipment drives the separation cup to rotate at high speed; Device B uses a peristaltic pump as a power source to transport the biological fluid from the liquid inlet channel to the hemispherical flow-collecting hole at the bottom of the inner core; under the action of centrifugal force, the biological fluid quickly climbs along the diversion channel to the annular baffle, and then quickly climbs along the waist-shaped hole and the drainage cone surface to the separation chamber, and the cell fluid gradually fills the separation chamber; When the biological fluid fills the separation chamber, the cells in the biological fluid are distributed on the outermost side of the separation chamber under the action of centrifugal force, while the waste fluid is located on the innermost side of the separation chamber; C With the continuous input of biological fluid, the waste liquid without cells on the innermost side of the separation chamber is squeezed out; when the biological fluid is completely input into the separation cup, the outer side of the separation chamber is cells and the inner side is waste liquid, and the total volume is the maximum dynamic volume of the separation cup; D. Start the air pump, push the piston, and cooperate with the gravity sensor, optical sensor and valve components on the separation equipment until a certain volume remains in the separation cup, and then stop pushing the piston; E The centrifuge is stopped and the air pump is started to pump air in the reverse direction to move the piston to the bottom. Under the action of gravity, all the biological fluid in the separation cup is collected at the hemispherical flow-collecting hole at the bottom of the inner core. Then the peristaltic pump is started to suck away the remaining biological fluid from the guide tube. F. A small amount of cleaning liquid is sucked in from the liquid outlet, and the centrifuge is started to clean the liquid outlet, the liquid outlet channel, the separation chamber and the guide channel; The peristaltic pump is then started again to draw the remaining cells from the liquid inlet channel into the product bag.

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