A cell preparation multi-connected cryogenic tube and a filling system and method

Abstract

The application relates to a cell preparation multi-connection cryopreservation tube and a filling system and method, which comprises a multi-connection cryopreservation tube 10 and a constant-pressure injection tube 20, the multi-connection cryopreservation tube is provided with a plurality of unit cryopreservation tubes 11, the unit cryopreservation tubes are connected in series through connecting tubes 12, the multi-connection cryopreservation tube is provided with a liquid injection tube 13 and an exhaust tube 14, the constant-pressure injection tube is provided with a liquid cavity 21 and a gas cavity 22, the liquid cavity is communicated with the liquid injection tube 13 of the multi-connection cryopreservation tube, the liquid cavity is also communicated with a cell bag 30, the gas cavity 22 is communicated with the exhaust tube 14 of the multi-connection cryopreservation tube, and the gas cavity is also communicated with the cell bag 30. The cell preparation is stored in the multi-connection cryopreservation tube, the cell preparation can be quickly and large-batch small-specification divided, and different-dose cell preparations can be conveniently taken as required. The constant-pressure injection tube system can realize the functions of automatic extraction, automatic filling and automatic recovery of cell preparations, the pipeline system is fully closed, the whole process of cell preparation filling is guaranteed to be sterile, and the safety of the cell preparation is protected.

Description

Technical Field

[0001] This invention relates to cell preparation cryopreservation tubes and filling devices, and more particularly to a cell preparation multi-cell cryopreservation tube and filling system and method. Background Technology

[0002] As a cutting-edge product of personalized medicine, cell therapy drugs rely heavily on the final step of their production process—formulation filling and cryopreservation—to ensure drug safety and efficacy. Cellular preparations typically require long-term storage in liquid nitrogen (-196°C). Currently, the main formulation containers used in the cell therapy industry include cryovials and cryopreservation bags. Traditional screw-cap cryovials are compact and easy to handle, suitable for dispensing small volumes of cell preparations. However, their screw cap structure relies on manual torque operation, making it difficult to precisely control the sealing force on automated filling lines, posing risks of leakage and external contamination. Furthermore, the tube openings often lack aseptic connections for disposable filling consumables, hindering fully enclosed automated filling. Cryopreservation bags are widely used for large-volume storage of autologous and allogeneic cell preparations, and they are easier to handle with fully enclosed aseptic connections and filling. However, due to their structural characteristics, cryopreservation bags are unsuitable for small-volume dispensing. Secondly, the bag and tubing design can lead to high levels of formulation residue during clinical use, resulting in significant product value loss.

[0003] Automated filling equipment is an effective means to ensure consistency, aseptic protection, and process scalability in cell therapy formulation dispensing. Although multiple brands of systems exist on the market, their basic process principles are similar: disposable sterile consumables are used to deliver the formulation in a completely closed path, a high-precision peristaltic pump is used as the power core for quantitative dispensing, and a programmable logic controller coordinates the entire process. Structurally, there are significant differences between filling cryovials and cryopreservation bags: cryovial filling often uses a rotary track for high-speed positioning, automatic capping and torque monitoring for opening and closing, and liquid addition via a filling needle inserted into the tube opening. These devices are often large and require a dedicated, fully sealed cleanroom or installation in a Class 100 cleanroom. Cryopreservation bag filling mainly achieves closed filling through tubing connection. Tubing can be manually connected on a clean bench or designed for automatic connection. After filling, the inlet tubing is heat-sealed and cut off, and air must be purged from the tube before filling to prevent breakage during cryopreservation. These modular designs collectively support the commercial production needs of cell therapy products.

[0004] The existing technologies have several drawbacks: traditional cryovials and cryopreservation bags have significant limitations in their application. Cryovials require opening the cap, posing a high risk of contamination. When freezing large-volume samples, several cryovials need to be used simultaneously, making the process cumbersome. Automated filling equipment is bulky, expensive, and has high installation requirements, making it unsuitable for dispensing cell preparations into cryovials. Furthermore, even with automated filling equipment, multiple cryovials still need to be handled separately during clinical use, increasing processing time and negatively impacting cell viability. Cryopreservation bags, on the other hand, cannot be made in very small sizes; the standard size is at least 10mL, making them unsuitable for small-volume dispensing. Currently, with the continuous development and upgrading of cryoprotectant products, high cell density cryopreservation can be achieved. In clinical use, diluting cell preparations with conventional injectable solutions such as physiological saline will significantly reduce the concentration of potentially sensitizing cryoprotectant components like DMSO infused into patients, further limiting the applicability of cryopreservation bags in the future. Summary of the Invention

[0005] The purpose of this invention is to provide a multi-cell cryopreservation tube filling system and method for cell preparations, making the cryopreservation and use of cell preparations more convenient and effective.

[0006] To achieve the above objectives, the technical solution of the present invention is: a cell preparation multi-cell cryopreservation tube, wherein the multi-cell cryopreservation tube 10 is provided with multiple unit cryopreservation tubes 11 and a multi-cell support 15, the multiple unit cryopreservation tubes 11 are arranged and installed on the multi-cell support 15, each unit cryopreservation tube 11 is provided with a unit input tube 111 and a unit output tube 112, the unit input tube 111 is connected to the inner cavity of the unit cryopreservation tube 11, the unit output tube 112 is inserted into the inner cavity of the unit cryopreservation tube 11, the unit input tube 111 of the unit cryopreservation tube at the first end is connected to an injection tube 13, the unit output tube 112 of the unit cryopreservation tube at the last end is connected to a discharge tube 14, and the unit input tubes 111 and unit output tubes 112 of two adjacent unit cryopreservation tubes are connected by a connecting tube 12.

[0007] Furthermore, a preferred unit cryopreservation tube structure is as follows: the unit cryopreservation tube 11 includes a cryopreservation tube body 113, a cryopreservation tube top cap 114, and a cryopreservation tube bottom cap 115. The cryopreservation tube top cap 114 is provided with the unit input tube 111 and the unit output tube 112. A sealing ring 116 is provided between the cryopreservation tube top cap 114 and the cryopreservation tube body 113. The bottom end of the cryopreservation tube body 113 is provided with a tube body bottom hole 117. The cryopreservation tube bottom cap 115 is provided with a bottom cap hole 118, which corresponds to the tube body bottom hole 117. A silicone plug 119 is provided between the bottom end of the cryopreservation tube body 113 and the cryopreservation tube bottom cap 115.

[0008] Furthermore, in order to achieve the arrangement and installation of multiple unit cryopreservation tubes, the multi-unit frame 15 is provided with multiple cryopreservation tube retaining rings 151, each cryopreservation tube retaining ring having a retaining ring opening 152, and a connecting strip 153 being provided between adjacent cryopreservation tube retaining rings 151.

[0009] Furthermore, in order to achieve structural versatility and adapt to cryopreservation racks, the height H of the multiple unit cryopreservation tubes 11 is equal.

[0010] A cell preparation multi-cell cryopreservation tube filling system includes the aforementioned multi-cell cryopreservation tube. The filling system is equipped with a constant pressure injection tube 20, which has a liquid chamber 21 and a gas chamber 22. The liquid chamber 21 is equipped with a liquid chamber piston 23, and the gas chamber 22 is equipped with a gas chamber piston 24. The liquid chamber piston 23 and the gas chamber piston 24 move synchronously. The liquid chamber 21 is connected to the liquid interface 31 of the cell bag 30 and the injection tube 13 of the multi-cell cryopreservation tube, respectively. The gas chamber 22 is connected to the gas interface 32 of the cell bag 30, and the discharge tube 14 of the multi-cell cryopreservation tube is connected to the liquid interface 31 of the cell bag 30.

[0011] Furthermore, in order to achieve synchronous movement of the liquid chamber piston and the gas chamber piston, the constant pressure injection tube 20 is vertically arranged, and a piston rod 27 is provided inside the constant pressure injection tube 20. The lower end of the piston rod 27 is provided with the liquid chamber piston 23, and the upper end of the piston rod 27 is provided with the gas chamber piston 24. A driving slider 28 is provided in the middle of the piston rod 27, and the tube wall of the constant pressure injection tube 20 is provided with a sliding groove 29. The driving slider 28 moves in the sliding groove 29, and the driving slider 28 drives the piston rod 27 to move.

[0012] Furthermore, a preferred tubing connection structure is as follows: the lower end of the constant pressure injection tube 20 is provided with a liquid interface 25, which is connected to the liquid chamber 21. The liquid interface 25 is connected to a first conduit 41. The first conduit 41 is connected to a second conduit 42 and a third conduit 43 via a tee. The second conduit 42 is connected to the injection tube 13 of the multi-cell cryopreservation tube. The discharge tube 14 of the multi-cell cryopreservation tube is connected to a fourth conduit 44. The third conduit 43 and the fourth conduit 44 are connected to a fifth conduit 45 via a tee. The fifth conduit 45 is connected to the liquid interface 31 of the cell bag 30. The upper end of the constant pressure injection tube 20 is provided with a gas interface 26, which is connected to the gas chamber 22. The gas interface 26 is connected to the gas interface 32 of the cell bag 30 via a sixth conduit 46.

[0013] Furthermore, to enable the constant pressure injection tubing and connecting tubing to form a movable installation structure, the first catheter 41, the second catheter 42, the third catheter 43, the fourth catheter 44, the fifth catheter 45, and the sixth catheter 46 are medical tubing. The first catheter 41 is fixedly connected to the liquid interface 25, the sixth catheter 46 is fixedly connected to the gas interface 26, the second catheter 42 is movably connected to the injection tube 13 of the multi-cell cryopreservation tube, the fourth catheter 44 is movably connected to the discharge tube 14 of the multi-cell cryopreservation tube, the fifth catheter 45 is movably connected to the liquid interface 31 of the cell bag 30, and the sixth catheter 46 is movably connected to the gas interface 32 of the cell bag 30.

[0014] Furthermore, in order to control the filling and recovery of cell preparations, the system is equipped with a first control valve 51, a second control valve 52, and a third control valve 53. The first control valve 51 controls the opening and closing of the third conduit 43, the second control valve 52 controls the opening and closing of the second conduit 42, and the third control valve 53 controls the opening and closing of the fourth conduit 44. The system is equipped with a first gas-liquid detector 61 and a second gas-liquid detector 62. The first conduit 41 passes through the first gas-liquid detector 61, and the fifth conduit 45 passes through the second gas-liquid detector 62.

[0015] A method for filling cell preparation multi-cell cryovials, comprising the cell preparation multi-cell cryovials and system as described in claims 1 to 9, characterized in that the method involves filling the cell preparation from the cell bag 30 into the multi-cell cryovial 10, and the method includes the following steps: a. In the initial state, a drainage volume 211 is reserved in the liquid chamber 21 of the constant pressure injection tube 20. The capacity of the drainage volume 211 is not less than the sum of the pipeline capacities of the first conduit 41, the second conduit 42, the fourth conduit 44, the fifth conduit 45 and the connecting tube 12 of the multi-unit cryopreservation tube. b. Connect the second conduit 42 to the injection tube 13 of the multi-cell cryopreservation tube, connect the fourth conduit 44 to the discharge tube 14 of the multi-cell cryopreservation tube, connect the fifth conduit 45 to the liquid interface 31 of the cell bag 30, and connect the sixth conduit 46 to the gas interface 32 of the cell bag 30. c. Open the first control valve 51 and close the second control valve 52 and the third control valve 53; d. Drive the piston rod 27 to move upward, and the cell preparation in the cell bag 30 enters the liquid chamber 21 through the third conduit 43 and the first conduit 41. The gas in the gas chamber 22 enters the cell bag 30 through the sixth conduit 46. When the first gas-liquid detector 61 detects gas passing through the first conduit 41, the piston rod 27 stops moving. e. Close the first control valve 51, and open the second control valve 52 and the third control valve 53; f. Drive the piston rod 27 downward, and the cell preparation in the liquid chamber 21 enters the multi-cell cryopreservation tube through the first conduit 41, the second conduit 42, and the injection tube 13 of the multi-cell cryopreservation tube, filling each unit cryopreservation tube 11 in turn. In the inner cavity of the unit cryopreservation tube 11, the lower end of the unit output tube 112 is the upper limit of the filling of the unit cryopreservation tube. When all the cell preparation in the liquid chamber 21 is output, the gas in the liquid chamber 21 enters the multi-cell cryopreservation tube 10. In the unit cryopreservation tube, the gas enters the connecting tube 12 from the gap between the lower end of the unit output tube 112 and the liquid surface of the cell preparation, and enters the cell bag 30 through the fourth conduit 44 and the fifth conduit 45. In this step, the gas in the cell bag 30 enters the gas chamber 22 through the sixth conduit 46. g. The filling operation ends when all the cell preparations and gases in the liquid chamber 21 have been discharged.

[0016] The beneficial effects of this invention are: The use of multi-cell cryopreservation tubes for preserving cell preparations allows for rapid, large-scale, small-specification dispensing of cell preparations and facilitates on-demand dispensing of different dosages. The constant-pressure injection tubing system enables automatic extraction, automatic filling, and automatic recovery of cell preparations. The fully enclosed tubing system ensures sterility throughout the cell preparation filling process, protecting the safety of the cell preparations.

[0017] The present invention will now be described in detail with reference to the accompanying drawings and embodiments. Attached Figure Description

[0018] Figure 1 This is a structural diagram of the multi-unit cryopreservation tube of the present invention; Figure 2 This is an exploded view of the structure of the multi-unit cryopreservation tube of the present invention; Figure 3 These are cross-sectional views and exploded views of the cryopreservation tube of this invention. Figure 4 This is a schematic diagram of the filling of the multi-cell cryopreservation tubes of the present invention, showing the cell preparation entering the multi-cell cryopreservation tubes; Figure 5 This is a schematic diagram of the filling of the multi-cell cryopreservation tube of the present invention. The cell preparation completely fills the unit cryopreservation tube, and the remaining cell preparation is discharged. Figure 6 This is a schematic diagram of the multi-cell cryopreservation tube filling process of the present invention. The cell preparation completely fills the unit cryopreservation tube, and the gas is discharged through the multi-cell cryopreservation tube. Figure 7 This is a schematic diagram of the system of the present invention; Figure 8 This is a structural diagram of the constant pressure injection tube of the present invention; Figure 9It is a schematic diagram of the assembly consisting of a constant pressure injection tube, a first catheter, a second catheter, a third catheter, a fourth catheter, a fifth catheter, and a sixth catheter; Figure 10 This is a structural diagram of the filling machine of the present invention; Figure 11 This is a schematic diagram showing the connection between the constant pressure injection tube, the multi-unit cryopreservation tube, and the cell bag on the filling machine of this invention; Figure 12 This is an operational schematic diagram of the present invention, showing the starting state of the filling operation; Figure 13 This is a schematic diagram of the operation of the present invention. The cell preparation in the cell bag is drawn into the liquid chamber of the constant pressure injection tube, and air is pushed into the cell bag by the gas. Figure 14 This is a schematic diagram of the operation of the present invention. The cell preparation in the liquid chamber is pushed into the multi-cell cryopreservation tube, and the air in the multi-cell cryopreservation tube is drawn into the gas chamber. Figure 15 This is a schematic diagram of the operation of the present invention, in which the remaining cell preparation is recycled into the cell bag; Figure 16 This is a schematic diagram of the operation of using the unit cryopreservation tube. Detailed Implementation Example 1:

[0019] like Figures 1 to 6 A multi-unit cryopreservation tube 10 is provided, comprising multiple unit cryopreservation tubes 11. In this embodiment, the multi-unit cryopreservation tube 10 comprises five unit cryopreservation tubes 11, which are connected by a multi-unit frame 15 and arranged sequentially in a row on the multi-unit frame 15. The multi-unit frame 15 is provided with multiple cryopreservation tube retaining rings 151, each with a retaining ring opening 152 for easy insertion of the unit cryopreservation tubes 11 into the multi-unit frame 15. A connecting strip 153 is provided between adjacent cryopreservation tube retaining rings 151.

[0020] The unit cryopreservation tube 11 includes a cryopreservation tube body 113, with a top cap 114 and a bottom cap 115 at each end, forming the inner cavity of the cryopreservation tube. The top cap 114 has a unit input pipe 111 and a unit output pipe 112. The unit input pipe 111 connects to the inner cavity of the cryopreservation tube at the top cap, and the unit output pipe 112 is inserted into the inner cavity of the unit cryopreservation tube 11, extending a certain length into the inner cavity and communicating with it. A sealing ring 116 is provided between the top cap 114 and the cryopreservation tube body 113. The top cap 114 and the cryopreservation tube body 113 are connected by a sealing adhesive or a threaded connection.

[0021] The cryopreservation tube body 113 has two tube body pin holes 117 at its bottom end, and the cryopreservation tube bottom cap 115 has two bottom cap pin holes 118, which correspond to the two tube body pin holes 117. A silicone plug 119 is provided between the bottom end of the cryopreservation tube body 113 and the cryopreservation tube bottom cap 115. The cryopreservation tube body 113 and the cryopreservation tube bottom cap 115 are connected by a sealing adhesive or a threaded connection.

[0022] Each cryopreservation unit 11, arranged at both ends, is equipped with an injection tube 13 and an outlet tube 14. The unit input tube 111 of the first cryopreservation unit is connected to the injection tube 13, and the unit output tube 112 of the last cryopreservation unit is connected to the outlet tube 14. The five cryopreservation units 11 are connected in series via connecting tubes 12, with the unit input tubes 111 and unit output tubes 112 of adjacent cryopreservation units 11 connected by connecting tubes 12. Thus, the five cryopreservation units form a series structure, with the unit input tubes 111 and unit output tubes 112 serving as interfaces at both ends of the multi-unit cryopreservation unit 10. The unit input tube 111 of each cryopreservation unit serves as the input interface for cell preparations, and the unit output tube 112 of each cryopreservation unit serves as an venting interface or an output interface for cell preparations.

[0023] The inlets of the injection tube 13 and the discharge tube 14 can be designed with external threads for use in sterile environments. Alternatively, the inlets of the injection tube 13 and the discharge tube 14 can be designed with silicone plugs for use in general environments. Protective caps can be fitted to the inlets of the injection tube 13 and the discharge tube 14.

[0024] like Figure 4 As shown, cell preparation 33 enters the inner cavity of the cryopreservation tube from the unit input tube 111 at the first end. As the cell preparation is injected, when the liquid level in the first unit cryopreservation tube 11 reaches the lower end of the unit output tube 112, the liquid level in the unit cryopreservation tube 11 will no longer rise due to the sealed space above the liquid level. The cell preparation then enters the connecting tube 12 through the unit output tube 112, and then enters the unit input tube 111 of the next unit cryopreservation tube, and finally enters the inner cavity of this unit cryopreservation tube 11. This process is repeated continuously until the inner cavities of multiple unit cryopreservation tubes 11 are fully filled.

[0025] After that, as Figure 5 As shown, if cell preparation 33 continues to enter the first unit cryopreservation tube, the cell preparation will be discharged from the discharge tube 14 through the unit output tube 112 of the last unit cryopreservation tube.

[0026] like Figure 6As shown, if gas 34 enters the first unit cryopreservation tube 11, the gas 34 will compress the cell preparation 33 inside the unit cryopreservation tube 11, creating a small gap H2 between the cell preparation 33 and the lower end of the unit output tube 112. The gas 34 will then pass through the connecting tube 12 through multiple unit cryopreservation tubes, sequentially discharging the cell preparation from each unit input tube 111, unit output tube 112, and connecting tube 12 through the discharge tube 14. This allows for the recovery of the cell preparation from the unit input tube 111, unit output tube 112, and connecting tube 12.

[0027] After being filled, the cryovial 11 will retain gas to prevent the cryovial from bursting due to the expansion of the cell preparation during freezing.

[0028] The height H (i.e., the length of the unit cryopreservation tube) of the five unit cryopreservation tubes is equal. Changing the height H of the unit cryopreservation tube can change the standard filling capacity of the unit cryopreservation tube.

[0029] The height (i.e., length) H of the five unit cryopreservation tubes 11 is equal. By changing the depth H1 of the unit output tube 112 inserted into the inner cavity of the unit cryopreservation tube 11, the capacity of the unit cryopreservation tube can be changed. Thus, by changing different cryopreservation tube top caps 114 and configuring different depths H1 of the unit output tube 11 inserted into the inner cavity of the unit cryopreservation tube 11, the filling capacity of the unit cryopreservation tube 11 can be controlled. Example 2:

[0030] like Figures 7 to 11 A cell preparation multi-cell cryopreservation tube filling system includes the multi-cell cryopreservation tube 10 described in Example 1. The filling system includes a constant-pressure injection tube 20, which is used to draw cell preparation from a cell bag 30 and fill the cell preparation into the multi-cell cryopreservation tube 10. The constant-pressure injection tube 20 is a cylindrical tube, vertically arranged, with a liquid chamber 21 at its lower part and a gas chamber 22 at its upper part. A liquid interface 25 is located at the lower end of the constant-pressure injection tube 20, connecting to the liquid chamber 21, and a gas interface 26 is located at the upper end of the constant-pressure injection tube 20, connecting to the gas chamber 22.

[0031] A liquid chamber piston 23 is provided in the liquid chamber 21, and the liquid chamber piston 23 moves within the liquid chamber 21. A gas chamber piston 24 is provided in the gas chamber 22, and the gas chamber piston 24 moves within the gas chamber 22. A piston rod 27 is provided in the constant pressure injection tube 20. The liquid chamber piston 23 is installed at the lower end of the piston rod 27, and the gas chamber piston 24 is installed at the upper end of the piston rod 27. A drive slider 28 is provided in the middle of the piston rod 27, and the drive slider 28 is fixed to the piston rod 27. The tube wall of the constant pressure injection tube 20 is provided with a sliding groove 29. The drive slider 28 moves in the sliding groove 29, and the drive slider 28 drives the piston rod 27 to move. The piston rod 27 drives the gas chamber piston 23 and the liquid chamber piston 24 to move synchronously.

[0032] The cell bag 30 has a liquid interface 31 at the lower end and a gas interface 32 at the upper end.

[0033] The connection between the multi-cell cryopreservation tube 10, the constant-pressure injection tube 20, and the cell bag 30 is as follows: the liquid chamber 21 of the constant-pressure injection tube is connected to the liquid interface 31 of the cell bag 30 and the injection tube 13 of the multi-cell cryopreservation tube, respectively; the discharge tube 14 of the multi-cell cryopreservation tube is connected to the liquid interface 31 of the cell bag 30; and the gas chamber 22 of the constant-pressure injection tube is connected to the gas interface 32 of the cell bag 30.

[0034] As a specific connection structure, the liquid interface 25 of the constant pressure injection tube is connected to the first conduit 41. The first conduit 41 is connected to the second conduit 42 and the third conduit 43 via a tee. The second conduit 42 is connected to the injection tube 13 of the multi-cell cryopreservation tube, and the third conduit 43 is connected to the liquid interface 31 of the cell bag 30. The discharge tube 14 of the multi-cell cryopreservation tube is connected to the fourth conduit 44. The fourth conduit 44 and the third conduit 43 are connected to the fifth conduit 45 via a tee. The fifth conduit 45 is connected to the liquid interface 31 of the cell bag. The gas interface 26 of the constant pressure injection tube is connected to the sixth conduit 46, and the sixth conduit 46 is connected to the gas interface 32 of the cell bag 30.

[0035] The first catheter 41, the second catheter 42, the third catheter 43, the fourth catheter 44, the fifth catheter 45, and the sixth catheter 46 are flexible tubes.

[0036] The first conduit 41 is fixedly connected to the liquid interface 25 of the constant pressure injection tube, and the sixth conduit 46 is fixedly connected to the gas interface 26 of the constant pressure injection tube. The constant pressure injection tube 20, the first conduit 41, the third conduit 43, the second conduit 42, the fourth conduit 44, the fifth conduit 45, and the sixth conduit 46 constitute a movable assembly, such as... Figure 9 As shown, it can be used as a disposable medical product to avoid cross-contamination.

[0037] In use, the fifth conduit 45 is movably connected to the liquid interface 31 of the cell bag 30, the sixth conduit 46 is movably connected to the gas interface 32 of the cell bag 30, the second conduit 42 is movably connected to the injection tube 13 of the multi-cell cryopreservation tube, and the fourth conduit 44 is movably connected to the discharge tube 14 of the multi-cell cryopreservation tube. The connectors for the fifth conduit 45 to the liquid interface 31 of the cell bag 30, the sixth conduit 46 to the gas interface 32 of the cell bag 30, the second conduit 42 to the injection tube 13 of the multi-cell cryopreservation tube, and the fourth conduit 44 to the discharge tube 14 of the multi-cell cryopreservation tube can be designed with internal threads (suitable for sealed cell containers) or long needles (suitable for open cell containers).

[0038] To control the filling process, the filling system is equipped with a first control valve 51, a second control valve 52, and a third control valve 53. The first control valve 51, the second control valve 52, and the third control valve 53 are electrically operated clamp valves.

[0039] The third conduit 43 is embedded in the first control valve 51, which controls the opening and closing of the third conduit 43. The second conduit 42 is embedded in the second control valve 52, which controls the opening and closing of the second conduit 42. The fourth conduit 44 is embedded in the third control valve 53, which controls the opening and closing of the fourth conduit 44.

[0040] The filling system is also equipped with a first gas-liquid detector 61 and a second gas-liquid detector 62, which are ultrasonic bubble detectors.

[0041] The first conduit 41 passes through the first gas-liquid detector 61, and the fifth conduit 45 passes through the second gas-liquid detector 62.

[0042] like Figure 10 , Figure 11 As shown, in order to achieve automatic filling, the filling system of this embodiment is provided with a filling machine 70. The constant pressure injection tube 20 is installed on the filling machine 70. The filling machine 70 is provided with a drive clamp 71 and a drive slide rail 72. The drive clamp 71 moves up and down along the drive slide rail 72. The drive clamp 71 is clamped on the drive slider 28 of the constant pressure injection tube, which drives the drive slider 28 to move up and down, thereby driving the air chamber piston 23 and the liquid chamber piston 24 to move up and down synchronously.

[0043] The filling machine 70 is equipped with an injection tube holder 77 and an injection tube fixing clamp 78. The lower end of the constant pressure injection tube 20 is placed on the injection tube holder 77, and the outer diameter of the constant pressure injection tube 20 is clamped on the injection tube fixing clamp 78.

[0044] The constant pressure injection tube 20, the first catheter 41, the second catheter 42, the third catheter 43, the fourth catheter 44, the fifth catheter 45, and the sixth catheter 46 can be movably installed on the filling machine 70 as disposable components to avoid cross-contamination of cell preparations.

[0045] The filling machine 70 is equipped with a cryopreservation box slot 76, in which multiple cryopreservation tubes 10 are placed, and the cryopreservation box 73 is placed in the cryopreservation box slot 76. The filling machine 70 is equipped with a cell bag hook 74, on which cell bags 30 are suspended. The filling machine is equipped with an operation panel 75 for controlling the filling of cell-based products. Example 3:

[0046] A method for filling cell preparation multi-cell cryovials is provided, which uses the cell preparation multi-cell cryovials and systems described in Examples 1 and 2 to achieve the filling of multi-cell cryovials.

[0047] After cell culture, cells are collected and washed automatically or manually. The harvested cells are then resuspended in cryopreservation solution in sealed cell bags (automatic harvesting method) or centrifuge tubes (manual operation method). This method allows cell preparations to be filled into multiple cryopreservation tubes 10. This embodiment demonstrates the dispensing of cell preparations within sealed cell bags (automatic harvesting method).

[0048] During operation, the sealed cell bag 30 is hung on the hook 74, the multi-cell cryopreservation tube 10 is inserted into the tray 73, and the constant pressure injection tube 20 is fixed on the filling machine.

[0049] like Figures 12 to 15 The filling steps of this formula include: a. such as Figure 12 As shown, in the initial state, a drainage volume 211 is reserved in the liquid chamber 21 of the constant pressure injection tube 20. The capacity of the drainage volume 211 is not less than the sum of the pipeline capacities of the first conduit 41, the second conduit 42, the fourth conduit 44, the fifth conduit 45 and the connecting tube 12 of the multi-unit cryopreservation tube.

[0050] b. According to the system described in Example 2, the second conduit 42 is connected to the injection tube 13 of the multi-cell cryopreservation tube, the fourth conduit 44 is connected to the discharge tube 14 of the multi-cell cryopreservation tube, the fifth conduit 45 is connected to the liquid interface 31 of the cell bag 30, and the sixth conduit 46 is connected to the gas interface 32 of the cell bag 30.

[0051] c. For example Figure 13 As shown, cell preparation extraction is performed by opening the first control valve 51 and closing the second control valve 52 and the third control valve 53.

[0052] d. Start the drive jack 71, drive the piston rod 27 to move upward, and the cell preparation in the cell bag 30 enters the liquid chamber 21 through the third conduit 43 and the first conduit 41. At this time, the first gas-liquid detector 61 will show the liquid flow in the first conduit 41; the gas in the gas chamber 22 enters the cell bag 30 through the sixth conduit 46, and the pressure balance is maintained in the entire pipeline system; when the first gas-liquid detector 61 detects the gas passing through the first conduit 41, it indicates that the cell preparation in the cell bag has been completely extracted, the piston rod 27 stops moving, and the extraction of cell preparation is completed.

[0053] e. such as Figure 14 As shown, cell preparation filling is performed by closing the first control valve 51 and opening the second control valve 52 and the third control valve 53.

[0054] f. Drive the piston rod 27 downwards, and the cell preparation in the liquid chamber 21 enters the multi-cell cryopreservation tube through the first conduit 41, the second conduit 42, and the injection tube 13 of the multi-cell cryopreservation tube, successively filling each unit cryopreservation tube 11. As described in Example 1, the lower end of the unit output tube 112 is the upper limit of the filling capacity of the unit cryopreservation tube in the inner cavity of the unit cryopreservation tube 11. When all unit cryopreservation tubes 11 are completely filled, the remaining cell preparation is discharged through the discharge tube 14 and returned to the cell bag, realizing the recovery of the remaining cell preparation. After all the cell preparation in the liquid chamber 21 has been discharged, the gas in the liquid chamber 21 enters the multi-unit cryopreservation tube 10. Within the unit cryopreservation tube, the gas enters the connecting tube 12 through the gap between the lower end of the unit output tube 112 and the surface of the cell preparation liquid, and then enters the cell bag 30 via the fourth conduit 44 and the fifth conduit 45. This discharges the cell preparation from each unit input tube 111, unit output tube 112, and connecting tube 12 through the discharge tube 14 and returns it to the cell bag 30, achieving further recovery of the cell preparation. In this step, the gas in the cell bag 30 enters the gas chamber 22 through the sixth conduit 46, maintaining pressure balance throughout the entire piping system.

[0055] g. Once all the cell preparation and gas in the liquid chamber 21 have been expelled (i.e., the liquid chamber piston 23 has moved to the lowest point of the constant pressure injection tube 20), the filling operation ends. If the second gas detector 62 detects liquid passing through the fifth conduit 45, the operation screen 75 will prompt the user to recover the remaining cell preparation.

[0056] The filling process described above can be operated through the control panel 75, and the filling machine 70 can control the process to proceed automatically.

[0057] Parameters such as cell container type, cell volume, cryovial specifications, and number of cryovials can be set through the control panel.

[0058] After the filling operation is completed, do not disconnect the pipeline connection directly. Use a handheld mobile heat sealer to heat seal the injection tube 13 and discharge tube 14 of the multi-unit cryopreservation tube 10, and then remove the multi-unit cryopreservation tube 10 for subsequent cryopreservation work.

[0059] When using multiple cryovials 10, the cell preparation can be taken from all or only a few of the unit cryovials 11, depending on the need. Use a handheld heat sealer to heat seal the connecting tube 12 at the end of the desired unit cryovial, and break it off at the connecting strip 153. The remaining unit cryovials are returned to the liquid nitrogen tank for continued freezing. Figure 16As shown, three cryovial units were used. The cryovial units were rapidly thawed at 37°C. Then, the syringe needle 81 and air filter needle 82 were inserted into the cryovial units through the bottom cap insertion hole 118, silicone stopper 119, and tube insertion hole 117, respectively. The cryovials were then inverted, and the cell preparation was aspirated from the cryovials using the syringe until only the first cryovial unit remained. The cryovials were then upright, and the cell preparation in the first cryovial unit was completely aspirated. The syringe was then removed and injected directly into the patient, or diluted with an appropriate injection solution for infusion. The air filter needle 82 is equipped with a filter 821 to prevent air contamination of the cell preparation in the cryovials.

[0060] This invention uses a fully enclosed, series-connected integrated cryopreservation tube, which can realize the series connection of different numbers of unit cryopreservation tubes and fill them through a unified inlet, greatly improving filling efficiency, reducing the risk of contamination, and making it easy to automate.

[0061] During the filling of multi-cell cryopreservation tubes, the cell preparations pass sequentially through the injection tube, unit input tube, unit output tube, connecting tube, and discharge tube. The depth of the unit output tube into the unit cryopreservation tube controls the filling volume, achieving completely consistent equal-volume dispensing. Relying on its own physical structure rather than on automated filling equipment to control the dispensing volume greatly increases the accuracy and stability of dispensing.

[0062] The multi-unit cryopreservation tubing uses heat-sealable medical-grade tubing, along with a sealing ring, silicone plug, and air filter structure, enabling a sealed operation throughout its entire lifecycle from filling to clinical use, eliminating the risk of contamination.

[0063] Each cryotube unit has the same dimensions and structure as industry-standard cryotubes and can be placed in a regular cryobox without the need for custom-made cryoboxes of special sizes.

[0064] The multi-unit cryopreservation tubes adopt a heat-sealed and breakable connection structure, which allows for the retrieval of any number of unit cryopreservation tubes without affecting the continued freezing of other unit cryopreservation tubes.

[0065] The automatic filling device uses a constant pressure injection pipeline system instead of a traditional peristaltic pump system. The filling process is not affected by factors such as liquid viscosity, density, and pressure, which greatly improves the mechanical stability of the process.

[0066] The synchronous movement of the liquid chamber piston and the gas chamber piston in the constant pressure injection tube ensures that the pressure in the entire pipeline system, including the cryopreservation tube and cell bag, remains constant throughout the filling process. This prevents excessive or insufficient local pressure caused by pressure differences at both ends when there are many tubes in series, or the cell bag from being sucked down.

[0067] The automatic filling device is controlled by a gas-liquid detector and an electric valve, which can realize the fully automatic liquid aspiration and filling process and the automatic recovery of excess cell preparations.

[0068] The pipeline system interface adopts optional structures such as screw threads, silicone plugs, and pins, which can be connected to various manual and automatic cell harvesting scenarios in the current cell therapy industry.

[0069] The advantages and effects of this invention include: It combines the flexibility of small-volume dispensing of cryopreservation tubes with the safety of sealed filling of cryopreservation bags. It can achieve one-time small-volume dispensing, high-density cryopreservation, on-demand use, and one-time extraction, minimizing contamination and reduction of cell viability caused by the operation process, and reducing the amount of cryopreservation solution infused into the patient. High-precision dispensing is achieved through the structure of the cryovials themselves, without relying on an automated filling device. Therefore, depending on user needs, an automated filling device can be omitted, and manual operation with a regular syringe can still achieve efficient and high-precision dispensing.

[0070] It can stably and reliably complete the extraction, dispensing, and recycling of cell preparations, perfectly connecting various manual and automatic cell harvesting scenarios in the cell therapy industry.

Claims

1. A cell preparation multiplex cryopreservation tube, characterized in that, The multi-unit cryopreservation tube (10) is provided with multiple unit cryopreservation tubes (11) and a multi-unit rack (15). The multiple unit cryopreservation tubes (11) are arranged and installed on the multi-unit rack (15). Each unit cryopreservation tube (11) is provided with a unit input tube (111) and a unit output tube (112). The unit input tube (111) is connected to the inner cavity of the unit cryopreservation tube (11). The unit output tube (112) of the unit cryopreservation tube at the first end is connected to the injection tube (13). The unit output tube (112) of the unit cryopreservation tube at the end is connected to the discharge tube (14). The unit input tube (111) and the unit output tube (112) of two adjacent unit cryopreservation tubes are connected by a connecting tube (12).

2. The cell preparation multiplex cryopreservation tube according to claim 1, characterized in that, The unit cryopreservation tube (11) includes a cryopreservation tube body (113), a cryopreservation tube top cap (114), and a cryopreservation tube bottom cap (115). The cryopreservation tube top cap (114) is provided with the unit input tube (111) and the unit output tube (112). A sealing ring (116) is provided between the cryopreservation tube top cap (114) and the cryopreservation tube body (113). The bottom end of the cryopreservation tube body (113) is provided with a tube body bottom hole (117). The cryopreservation tube bottom cap (115) is provided with a bottom cap hole (118). The bottom cap hole (118) corresponds to the tube body bottom hole (117). A silicone plug (119) is provided between the bottom end of the cryopreservation tube body (113) and the cryopreservation tube bottom cap (115).

3. The cell preparation multiplex cryopreservation tube according to claim 1, characterized in that, The multi-unit rack (15) is provided with multiple cryopreservation tube clamps (151), each cryopreservation tube clamp has a clamp opening (152), and a connecting strip (153) is provided between adjacent cryopreservation tube clamps (151).

4. The cell preparation multiplex cryopreservation tube according to claim 1, characterized in that, The height (H) of the multiple cryopreservation tubes (11) is equal.

5. A cell preparation multi-cell cryopreservation tube filling system, comprising the multi-cell cryopreservation tubes as described in claims 1 to 4, characterized in that, The filling system is provided with a constant pressure injection tube (20), which is provided with a liquid chamber (21) and a gas chamber (22). The liquid chamber (21) is provided with a liquid chamber piston (23), and the gas chamber (22) is provided with a gas chamber piston (24). The liquid chamber piston (23) and the gas chamber piston (24) move synchronously. The liquid chamber (21) is connected to the liquid interface (31) of the cell bag (30) and the injection tube (13) of the multi-cell cryopreservation tube, respectively. The gas chamber (22) is connected to the gas interface (32) of the cell bag (30), and the discharge tube (14) of the multi-cell cryopreservation tube is connected to the liquid interface (31) of the cell bag (30).

6. The cell preparation multi-cell cryopreservation tube filling system according to claim 5, characterized in that, The constant pressure injection tube (20) is vertically arranged, and a piston rod (27) is provided inside the constant pressure injection tube (20). The lower end of the piston rod (27) is provided with the liquid chamber piston (23), and the upper part of the piston rod (27) is provided with the gas chamber piston (24). The middle part of the piston rod (27) is provided with a driving slider (28). The tube wall of the constant pressure injection tube (20) is provided with a sliding groove (29). The driving slider (28) moves in the sliding groove (29) and drives the piston rod (27) to move.

7. The cell preparation multi-cell cryopreservation tube filling system according to claim 5, characterized in that, The lower end of the constant pressure injection tube (20) is provided with a liquid interface (25), which is connected to the liquid chamber (21). The liquid interface (25) is connected to the first conduit (41). The first conduit (41) is connected to the second conduit (42) and the third conduit (43) respectively through a tee. The second conduit (42) is connected to the injection tube (13) of the multi-cell cryopreservation tube. The discharge tube (14) of the multi-cell cryopreservation tube is connected to the fourth conduit (44). The third conduit (43) and the fourth conduit (44) are connected to the fifth conduit (45) through a tee. The fifth conduit (45) is connected to the liquid interface (31) of the cell bag (30). The upper end of the constant pressure injection tube (20) is provided with a gas interface (26), which is connected to the gas chamber (22). The gas interface (26) is connected to the gas interface (32) of the cell bag (30) through a sixth conduit (46).

8. The cell preparation multi-cell cryopreservation tube filling system according to claim 7, characterized in that, The first catheter (41), the second catheter (42), the third catheter (43), the fourth catheter (44), the fifth catheter (45), and the sixth catheter (46) are medical tubing. The first catheter (41) is fixedly connected to the liquid interface (25), the sixth catheter (46) is fixedly connected to the gas interface (26), the second catheter (42) is movably connected to the injection tube (13) of the multi-cell cryopreservation tube, the fourth catheter (44) is movably connected to the discharge tube (14) of the multi-cell cryopreservation tube, the fifth catheter (45) is movably connected to the liquid interface (31) of the cell bag (30), and the sixth catheter (46) is movably connected to the gas interface (32) of the cell bag (30).

9. The cell preparation multi-cell cryopreservation tube filling system according to claim 7, characterized in that, The system is provided with a first control valve (51), a second control valve (52) and a third control valve (53). The first control valve (51) controls the opening and closing of the third conduit (43), the second control valve (52) controls the opening and closing of the second conduit (42), and the third control valve (53) controls the opening and closing of the fourth conduit (44). The system is provided with a first gas-liquid detector (61) and a second gas-liquid detector (62). The first conduit (41) passes through the first gas-liquid detector (61), and the fifth conduit (45) passes through the second gas-liquid detector (62).

10. A method for filling cell preparation multiple cryovials, comprising the cell preparation multiple cryovials and system as described in claims 1 to 9, characterized in that, The method involves filling the cell preparation from the cell bag (30) into the multi-cell cryovial (10), and the steps of the method include: a. In the initial state, a drainage volume (211) is reserved in the liquid chamber (21) of the constant pressure injection tube (20), and the capacity of the drainage volume (211) is not less than the sum of the pipeline capacities of the first conduit (41), the second conduit (42), the fourth conduit (44), the fifth conduit (45) and the connecting tube (12) of the multi-unit cryopreservation tube; b. Connect the second conduit (42) to the injection tube (13) of the multi-cell cryopreservation tube, connect the fourth conduit (44) to the discharge tube (14) of the multi-cell cryopreservation tube, connect the fifth conduit (45) to the liquid interface (31) of the cell bag (30), and connect the sixth conduit (46) to the gas interface (32) of the cell bag (30). c. Open the first control valve (51), and close the second control valve (52) and the third control valve (53); d. Drive the piston rod (27) to move upward, and the cell preparation in the cell bag (30) enters the liquid chamber (21) through the third conduit (43) and the first conduit (41). The gas in the gas chamber (22) enters the cell bag (30) through the sixth conduit (46). When the first gas-liquid detector (61) detects that gas is passing through the first conduit (41), the piston rod (27) stops moving. e. Close the first control valve (51) and open the second control valve (52) and the third control valve (53). f. Drive the piston rod (27) downward, and the cell preparation in the liquid chamber (21) enters the multi-cell cryopreservation tube through the first conduit (41), the second conduit (42), and the injection tube (13) of the multi-cell cryopreservation tube, filling each unit cryopreservation tube (11) in turn. In the inner cavity of the unit cryopreservation tube (11), the lower end of the unit output tube (112) is the upper limit of the filling of the unit cryopreservation tube. When all the cell preparation in the liquid chamber (21) is output, the gas in the liquid chamber (21) enters the multi-cell cryopreservation tube (10). In the unit cryopreservation tube, the gas enters the connecting tube (12) from the gap between the lower end of the unit output tube (112) and the liquid surface of the cell preparation, and enters the cell bag (30) through the fourth conduit (44) and the fifth conduit (45). In this step, the gas in the cell bag (30) enters the gas chamber (22) through the sixth conduit (46). g. The filling operation ends when all the cell preparations and gases in the liquid chamber (21) have been discharged.

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