Cell subpackaging method

Abstract

The invention provides a cell subpackaging method, and relates to the technical field of cell subpackaging. The subpackaging method comprises the following steps: communicating the subpackaging bag with a pre-filling bag in which pre-filling liquid is stored through a main pipeline; opening a corresponding pipeline electromagnetic valve on the subpackaging bag, and closing the pipeline electromagnetic valve after the peristaltic pump forwards rotates to pump the pre-filled liquid into the subpackaging bag; a corresponding pipeline electromagnetic valve on the split charging bag is opened, the peristaltic pump rotates reversely to pump the pre-filling liquid in the split charging bag into the pre-filling bag, and then the pipeline electromagnetic valve is closed; after being disconnected from the pre-filling bag, the main pipeline is connected to the storage bag in which the cell suspension is stored, the corresponding pipeline electromagnetic valve on the sub-packaging bag is opened, and after the peristaltic pump rotates forwards to pump the cell suspension into the sub-packaging bag, the pipeline electromagnetic valve is closed. The method can be applied to subpackaging operation of the cell preparation, air in the subpackaging bag can be removed, labor force can be reduced, the pollution risk is low, and the automation degree is high.

Description

Technical Field

[0001] The invention relates to the technical field of cell packaging, and in particular to a cell packaging method. Background Art

[0002] The production and manufacturing of clinical cell preparations is a complex process that uses a variety of technical means and is designed across multiple steps in different geographical locations. Any errors in production control, storage, or logistics transportation will cause cell damage and directly affect the final patient treatment efficiency. Among them, the packaging of cell preparations is a key step and starting step in cell production, and packaging will have a significant impact on the quality of cell products.

[0003] However, the current cell preparation packaging is mostly done manually, which makes it difficult to remove air and other gases from the packaging bags. After the cell preparations are packaged, some of the air remaining in the packaging bags will cause irreversible damage to the cells. Therefore, it is necessary to urgently propose a solution to improve the above problem. Summary of the invention

[0004] The purpose of the present invention is to provide a cell packaging method that can be applied to the packaging operation of cell preparations, which can not only remove the air in the packaging bag, but also reduce labor, has a low risk of contamination and a high degree of automation.

[0005] The present invention provides a cell packaging method, which adopts the following technical solution and comprises the following steps:

[0006] The sub-packaging bag with a capacity of V is connected to the pre-filling bag storing the pre-filling liquid through a main pipeline, and the connection point between the main pipeline and the sub-packaging bag is located at the top of the sub-packaging bag;

[0007] Open the corresponding pipeline solenoid valve on the sub-packaging bag, and the peristaltic pump rotates forward to 1 After the volume of pre-filling liquid is pumped into the dispensing bag, the pipeline solenoid valve is closed; V 1 : V = 1 / 5-3 / 5;

[0008] Open the corresponding pipeline solenoid valve on the sub-packaging bag, and the peristaltic pump reverses to transfer the V 2 After the volume of priming liquid is pumped into the priming bag, the pipeline solenoid valve is closed; among them, V 2 :V 1 =8 / 10-9 / 10;

[0009] The main pipeline is disconnected from the pre-filled bag and connected to a storage bag storing a cell suspension, the corresponding pipeline solenoid valve on the sub-packaging bag is opened, the peristaltic pump rotates forward to pump the cell suspension into the sub-packaging bag, and then the pipeline solenoid valve is closed; wherein the cell suspension is composed of cells and a resuspension medium, and the resuspension medium is of the same type as the pre-filled liquid.

[0010] The beneficial effect of the cell packaging method proposed by the present invention is that when the cell suspension is packaged, a prefill bag storing a prefill liquid is connected to a packaging bag in advance, and a peristaltic pump pumps the prefill liquid into the packaging bag, and the volume V of the prefill liquid pumped into the packaging bag is 1 The ratio of the capacity V of the sub-packaging bag is 1 / 5-3 / 5. Since the connection between the sub-packaging bag and the main pipeline is located at the top of the sub-packaging bag, the pre-filling liquid is located at the bottom of the sub-packaging bag. When the peristaltic pump reverses to pump the pre-filling liquid in the sub-packaging bag into the pre-filling bag, since there is gas at the top of the sub-packaging bag, the peristaltic pump first pumps the gas in the sub-packaging bag into the pre-filling bag. When the peristaltic pump pumps the V 2 When the volume of priming liquid is pumped into the priming bag, all the gas in the sub-packaging bag can be pumped into the priming bag, so that only the priming liquid remains in the sub-packaging bag. The peristaltic pump then rotates forward to pump the cell suspension into the sub-packaging bag, so that the cell suspension does not come into contact with gases such as air in the sub-packaging bag, and the resuspension medium in the cell suspension is of the same type as the priming liquid, that is, the priming liquid remaining in the sub-packaging bag will not affect the cells in the cell suspension.

[0011] Optionally, the step of connecting the sub-packaging bag with a capacity of V to the pre-filling bag storing the pre-filling liquid through the main pipeline includes: connecting N sub-packaging bags with a capacity of V to the pre-filling bag storing the pre-filling liquid through the main pipeline, wherein N is a positive integer greater than or equal to 2. After the N sub-packaging bags are connected to the main pipeline, the N sub-packaging bags can be pre-filled and vented, and after venting, the N sub-packaging bags can be sub-packed with the cell suspension, which can improve the packaging efficiency.

[0012] Optionally, the volume of the pre-filled liquid in the pre-filled bag is greater than N multiplied by V, so that there will be no shortage of pre-filled liquid when the pre-filled liquid is pumped into the sub-packaging bag.

[0013] Optionally, the step of opening the corresponding pipeline solenoid valve on the sub-packaging bag, and the peristaltic pump rotates forward to pump V1 volume of pre-filling liquid into the sub-packaging bag, and then closing the pipeline solenoid valve includes: the corresponding pipeline solenoid valves on N sub-packaging bags are opened and closed in sequence, and when the pipeline solenoid valve is opened but not closed, the peristaltic pump rotates forward to pump V1 volume of pre-filling liquid into the sub-packaging bag. 1 The volume of pre-filling liquid is pumped into the sub-packaging bag until all N sub-packaging bags are filled with pre-filling liquid. The pipeline solenoid valves on the N sub-packaging bags are opened and closed in sequence, so that the peristaltic pump can pump the pre-filling liquid into each sub-packaging bag when working.

[0014] Optionally, the corresponding pipeline solenoid valve on the sub-packaging bag is opened, and the peristaltic pump is reversed to transfer V 2 After the volume of priming liquid is pumped into the priming bag, the pipeline solenoid valve is closed. This step includes: the corresponding pipeline solenoid valves on the N sub-packaging bags are opened and closed in sequence. When the pipeline solenoid valve is opened but not closed, the peristaltic pump reverses to pump V in the sub-packaging bag. 2Volume of priming liquid is pumped into the priming bag until N sub-packaging bags have V 2 The volume of priming liquid is pumped into the priming bag. The peristaltic pump reverses to extract the liquid from the sub-packaging bag, and because the pipeline solenoid valves on the N sub-packaging bags are opened and closed in sequence, the priming liquid in the N sub-packaging bags can be extracted into the priming bag.

[0015] Optionally, the step of opening the corresponding pipeline solenoid valve on the sub-packaging bag, rotating the peristaltic pump forward to pump the cell suspension into the sub-packaging bag, and then closing the pipeline solenoid valve includes: the corresponding pipeline solenoid valves on N sub-packaging bags are opened and closed in sequence, and when the pipeline solenoid valve is opened but not closed yet, the peristaltic pump rotates forward to pump the cell suspension into the sub-packaging bag until the N sub-packaging bags are filled with cell suspension.

[0016] Optionally, the step of disconnecting the main pipeline from the pre-filled bag and connecting it to the storage bag storing the cell suspension, opening the corresponding pipeline solenoid valve on the sub-packaging bag, and closing the pipeline solenoid valve after the peristaltic pump rotates forward to pump the cell suspension into the sub-packaging bag, also includes: placing the storage bag on the mixing component, and the mixing component squeezes the bottom of the storage bag by 1-2 cm at a frequency of 60-70 times / min. When the peristaltic pump pumps the cell suspension in the storage bag into the sub-packaging bag, the mixing component squeezes the storage bag to mix the cell suspension in the storage bag evenly, avoiding the sedimentation of the cells in the cell suspension, so that the cells in the cell suspension pumped out by the peristaltic pump are evenly distributed.

[0017] Optionally, the peristaltic pump can be turned forward to 1 When the volume of pre-filling liquid is pumped into the dispensing bag, the forward speed of the peristaltic pump is 20-30rpm; the peristaltic pump is reversed to pump the V 2 When the volume of pre-filling solution is pumped into the pre-filling bag, the reverse speed of the peristaltic pump is 20-30rpm; when the peristaltic pump is rotated forward to pump the cell suspension into the filling bag, the forward speed of the peristaltic pump is 20-30rpm.

[0018] Optionally, the step of disconnecting the main pipeline from the pre-filled bag and then connecting it to the storage bag storing the cell suspension, opening the corresponding pipeline solenoid valve on the sub-packaging bag, and closing the pipeline solenoid valve after the peristaltic pump rotates forward to pump the cell suspension into the sub-packaging bag, further includes: a liquid flow meter provided on the main pipeline counts the volume of the cell suspension flowing into the sub-packaging bag. Thus, the volume of the cell suspension pumped into the sub-packaging bag is controlled, which is conducive to controlling the consistency of the volume of the cell suspension in the sub-packaging bag.

[0019] Optionally, the step of disconnecting the main pipeline from the pre-filled bag and connecting it to a storage bag storing a cell suspension, opening the corresponding pipeline solenoid valve on the sub-packaging bag, and rotating the peristaltic pump forward to pump the cell suspension into the sub-packaging bag, and then closing the pipeline solenoid valve, also includes: a bubble monitor set on the main pipeline to monitor whether there is gas passing through the main pipeline. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 is a flow chart of the steps of the cell packaging method in an embodiment of the present invention;

[0021] Figure 2 is a flowchart of the specific steps of step S2 in an embodiment of the present invention;

[0022] Figure 3 is a flowchart of the specific steps of step S3 in an embodiment of the present invention;

[0023] Figure 4 It is a specific step flow chart of step S4 in an embodiment of the present invention. DETAILED DESCRIPTION

[0024] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. Unless otherwise defined, the technical terms or scientific terms used herein should be understood by people with general skills in the field to which the present invention belongs. "Including" and similar words used in this article mean that the elements or objects appearing before the word include the elements or objects listed after the word and their equivalents, without excluding other elements or objects.

[0025] An embodiment of the present invention provides a cell packaging method.

[0026] See also Figure 1 , a cell packaging method comprising the following steps:

[0027] S1. Pre-filling pipeline connection: connect the sub-packaging bag with a capacity of V to the pre-filling bag storing the pre-filling liquid through the main pipeline, and the connection point between the main pipeline and the sub-packaging bag is located at the top of the sub-packaging bag;

[0028] S2. Pre-filling liquid: Open the corresponding pipeline solenoid valve on the sub-packaging bag, and the peristaltic pump rotates forward to V 1 After the volume of pre-filling liquid is pumped into the dispensing bag, the pipeline solenoid valve is closed; among them, V 1 : V = 1 / 5-3 / 5;

[0029] S3, exhaust of the sub-packaging bag: open the corresponding pipeline solenoid valve on the sub-packaging bag, and the peristaltic pump reverses to pump V2 volume of pre-filling liquid in the sub-packaging bag into the pre-filling bag, and then the pipeline solenoid valve is closed; wherein, V 2 :V 1 =8 / 10-9 / 10;

[0030] S4. Cell suspension subpackaging: disconnect the main pipeline from the pre-filled bag and connect it to the storage bag containing the cell suspension, open the corresponding pipeline solenoid valve on the sub-packaging bag, and after the peristaltic pump rotates forward to pump the cell suspension into the sub-packaging bag, the pipeline solenoid valve is closed; wherein, the cell suspension consists of cells and resuspension medium, and the resuspension medium is of the same type as the pre-filled liquid.

[0031] In some embodiments, the nominal capacity of the dispensing bag is one of 100 mL, 50 mL, 20 mL and 10 mL.

[0032] In some embodiments, the inner cavity volume of the main pipeline is 40 mL, that is, at most 40 mL of the priming solution or cell suspension remains in the main pipeline.

[0033] In some embodiments, the main pipeline is a disposable pipeline, and the main pipeline is replaced after completing a subpackaging operation.

[0034] In some embodiments, the process of executing step S1 includes: connecting N sub-packaging bags with a capacity of V to a pre-filling bag storing a pre-filling liquid through a main pipeline, and N is a positive integer greater than or equal to 2. The multiple sub-packaging bags are connected to the main pipeline, so that the pre-filling liquid can be pumped into the multiple sub-packaging bags, thereby improving the overall use efficiency.

[0035] In some embodiments, the process of executing step S1 includes: connecting N sub-packaging bags to a pre-filling bag storing pre-filling liquid through a main pipeline, where N is a positive integer greater than or equal to 2, and the capacities of the N sub-packaging bags can be the same or different.

[0036] In some embodiments, the volume of the pre-filled liquid in the pre-filled bag is greater than N times V, that is, the volume of the pre-filled liquid in the pre-filled bag is greater than the total capacity of N sub-packaging bags, so that the pre-filled liquid can be distributed to at least N sub-packaging bags, thereby completing the pre-filling operation of N sub-packaging bags with the pre-filled liquid.

[0037] In some embodiments, when N sub-packaging bags are connected to a pre-filling bag storing a pre-filling liquid through a main pipeline, executing step S2 includes: the pipeline solenoid valves on the N sub-packaging bags are opened and closed at one time, and when the pipeline solenoid valves are opened but not closed, the peristaltic pump rotates forward to turn V 1 The volume of priming liquid is pumped into the sub-filling bags until N sub-filling bags are filled with priming liquid.

[0038] In some embodiments, when N sub-packaging bags are connected to a pre-filled bag storing pre-filled liquid through a main pipeline, executing step S2 includes: the pipeline solenoid valves on the N sub-packaging bags are opened and closed in sequence, and when the pipeline solenoid valves are opened but not closed yet, the peristaltic pump rotates forward to pump the pre-filled liquid into the sub-packaging bags until all the N sub-packaging bags contain pre-filled liquid, and the volumes of the pre-filled liquid in the N sub-packaging bags may be the same or different, but the volume of the pre-filled liquid in the sub-packaging bags is at least 1 / 3 of the capacity of the pre-filled bags.

[0039] In some embodiments, see Figure 2 , when the four sub-packaging bags are connected to the pre-filling bag storing the pre-filling liquid through the main pipeline, and the four sub-packaging bags are respectively the first sub-packaging bag, the second sub-packaging bag, the third sub-packaging bag and the fourth sub-packaging bag in the direction away from the pre-filling bag, the pipeline solenoid valve corresponding to the first sub-packaging bag is the first pipeline solenoid valve, the pipeline solenoid valve corresponding to the second sub-packaging bag is the second pipeline solenoid valve, the pipeline solenoid valve corresponding to the third sub-packaging bag is the third pipeline solenoid valve, and the pipeline solenoid valve corresponding to the fourth sub-packaging bag is the fourth pipeline solenoid valve, executing step S2 includes:

[0040] S21, open the first pipeline solenoid valve, the second pipeline solenoid valve, the third pipeline solenoid valve and the fourth pipeline solenoid valve remain closed, and the peristaltic pump rotates forward to turn V 1 After the volume of pre-filling liquid is pumped into the first sub-packaging bag, the first pipeline solenoid valve and the peristaltic pump are closed; wherein, before the pre-filling liquid is pumped into the first sub-packaging bag, the pre-filling liquid pre-fills the main pipeline, and the gas in the main pipeline is discharged into the first sub-packaging bag;

[0041] S22, open the second pipeline solenoid valve, the first pipeline solenoid valve, the third pipeline solenoid valve and the fourth pipeline solenoid valve remain closed, and the peristaltic pump rotates forward to turn V 1 After the volume of pre-filling liquid is pumped into the second filling bag, the second pipeline solenoid valve and peristaltic pump are closed;

[0042] S23, open the third pipeline solenoid valve, the first pipeline solenoid valve, the second pipeline solenoid valve and the fourth pipeline solenoid valve remain closed, and the peristaltic pump rotates forward to turn V 1 After the volume of pre-filling liquid is pumped into the third filling bag, the solenoid valve and peristaltic pump of the third pipeline are closed;

[0043] S24, open the fourth pipeline solenoid valve, the first pipeline solenoid valve, the second pipeline solenoid valve and the third pipeline solenoid valve remain closed, and the peristaltic pump rotates forward to turn V 1 After the volume of priming liquid is pumped into the fourth filling bag, the fourth pipeline solenoid valve and the peristaltic pump are closed.

[0044] In some embodiments, when N sub-packaging bags are connected to the pre-filling bag storing the pre-filling liquid through the main pipeline, the execution of step S3 includes: the corresponding pipeline solenoid valves on the N sub-packaging bags are opened and closed in sequence, and when the pipeline solenoid valve is opened but not closed, the peristaltic pump is reversed to transfer the V in the sub-packaging bag to the pre-filling bag. 2 Volume of priming liquid is pumped into the priming bag until N sub-packaging bags have V 2 Pump volume of priming solution into the priming bag.

[0045] In some embodiments, when N sub-packaging bags are connected to a pre-filling bag storing pre-filling liquid through a main pipeline, executing step S3 includes: the corresponding pipeline solenoid valves on the N sub-packaging bags are opened and closed in sequence, and when the pipeline solenoid valves are opened but not closed, the peristaltic pump is reversed to pump the pre-filling liquid in the sub-packaging bag into the pre-filling bag, until the pre-filling liquid in the N sub-packaging bags is pumped into the pre-filling bag. The volumes of the pre-filling liquid pumped into the pre-filling bag in the N sub-packaging bags may be the same or different, but at least part of the pre-filling liquid in the N sub-packaging bags is pumped into the pre-filling bag.

[0046] In some embodiments, see Figure 3 , when the four sub-packaging bags are connected to the pre-filling bag storing the pre-filling liquid through the main pipeline, and the four sub-packaging bags are respectively the first sub-packaging bag, the second sub-packaging bag, the third sub-packaging bag and the fourth sub-packaging bag in the direction away from the pre-filling bag, the pipeline solenoid valve corresponding to the first sub-packaging bag is the first pipeline solenoid valve, the pipeline solenoid valve corresponding to the second sub-packaging bag is the second pipeline solenoid valve, the pipeline solenoid valve corresponding to the third sub-packaging bag is the third pipeline solenoid valve, and the pipeline solenoid valve corresponding to the fourth sub-packaging bag is the fourth pipeline solenoid valve, executing step S3 includes:

[0047] S31, open the first pipeline solenoid valve, keep the second pipeline solenoid valve, the third pipeline solenoid valve and the fourth pipeline solenoid valve closed, and reverse the peristaltic pump to transfer V 2 After the volume of priming liquid is pumped into the priming bag, the first pipeline solenoid valve and the peristaltic pump are closed;

[0048] S32, open the second pipeline solenoid valve, the first pipeline solenoid valve, the third pipeline solenoid valve and the fourth pipeline solenoid valve remain closed, and the peristaltic pump reverses to transfer V 2 After the volume of priming liquid is pumped into the priming bag, the second pipeline solenoid valve and the peristaltic pump are closed;

[0049] S33, open the solenoid valve of the third pipeline, keep the solenoid valve of the first pipeline, the solenoid valve of the second pipeline and the solenoid valve of the fourth pipeline closed, and reverse the peristaltic pump to transfer the V 2 After the volume of priming liquid is pumped into the priming bag, the solenoid valve and peristaltic pump of the third pipeline are closed;

[0050] S34, open the solenoid valve of the fourth pipeline, keep the solenoid valve of the first pipeline, the solenoid valve of the second pipeline and the solenoid valve of the third pipeline closed, and reverse the peristaltic pump to transfer the V 2 After the volume of priming liquid is pumped into the priming bag, the fourth pipeline solenoid valve and the peristaltic pump are closed.

[0051] In some embodiments, when N sub-packaging bags are connected to a pre-filled bag storing pre-filled liquid through a main pipeline, executing step S4 includes: the corresponding pipeline solenoid valves on the N sub-packaging bags are opened and closed in sequence, and when the pipeline solenoid valves are opened but not closed, the peristaltic pump rotates forward to pump the cell suspension into the sub-packaging bags until the cell suspension is stored in the N sub-packaging bags.

[0052] In some embodiments, when the N sub-packaging bags are connected to the pre-filling bag storing the pre-filling solution through a main pipeline, when step S4 is executed, the volumes of the cell suspension pumped into the N sub-packaging bags are the same.

[0053] In some embodiments, see Figure 4 , when the four sub-packaging bags are connected to the pre-filling bag storing the pre-filling liquid through the main pipeline, and the four sub-packaging bags are respectively the first sub-packaging bag, the second sub-packaging bag, the third sub-packaging bag and the fourth sub-packaging bag, the pipeline solenoid valve corresponding to the first sub-packaging bag is the first pipeline solenoid valve, the pipeline solenoid valve corresponding to the second sub-packaging bag is the second pipeline solenoid valve, the pipeline solenoid valve corresponding to the third sub-packaging bag is the third pipeline solenoid valve, and the pipeline solenoid valve corresponding to the fourth sub-packaging bag is the fourth pipeline solenoid valve, executing step S4 includes:

[0054] S41, after disconnecting the main pipeline from the pre-filled bag, connect the main pipeline to the storage bag storing the cell suspension;

[0055] S42, the first pipeline solenoid valve is opened, the second pipeline solenoid valve, the third pipeline solenoid valve and the fourth pipeline solenoid valve remain closed, the peristaltic pump rotates forward to pump the cell suspension in the storage bag into the first sub-packaging bag, and then the first pipeline solenoid valve and the peristaltic pump are closed;

[0056] S43, the second pipeline solenoid valve is opened, the first pipeline solenoid valve, the third pipeline solenoid valve and the fourth pipeline solenoid valve remain closed, the peristaltic pump rotates forward to pump the cell suspension in the storage bag into the second filling bag, and then the second pipeline solenoid valve and the peristaltic pump are closed;

[0057] S44, the solenoid valve of the third pipeline is opened, the solenoid valve of the first pipeline, the solenoid valve of the second pipeline and the solenoid valve of the fourth pipeline remain closed, the peristaltic pump rotates forward to pump the cell suspension in the storage bag into the third filling bag, and then the solenoid valve of the third pipeline and the peristaltic pump are closed;

[0058] S45, open the solenoid valve of the fourth pipeline, keep the solenoid valve of the first pipeline, the solenoid valve of the second pipeline and the solenoid valve of the third pipeline closed, and after the peristaltic pump rotates forward to pump the cell suspension in the storage bag into the fourth filling bag, the solenoid valve of the fourth pipeline and the peristaltic pump are closed.

[0059] In some embodiments, when the N sub-packaging bags are connected to the pre-filling bag storing the pre-filling solution through a main pipeline, when step S4 is executed, the volume of the cell suspension in the storage bag is greater than the amount to be sub-packed in the N sub-packaging bags.

[0060] In some embodiments, when N sub-packaging bags are connected to a pre-filled bag storing pre-filled liquid through a main pipeline, when step S4 is executed, when the volume of the cell suspension in the storage bag is less than the amount to be filled in the N sub-packaging bags, the following steps are executed: the storage bag is connected to the expansion bag using a connecting tube, wherein the expansion bag stores the cell suspension, and another peristaltic pump at the connecting tube rotates forward to pump the cell suspension in the expansion bag into the storage bag, so as to replenish the cell suspension in the storage bag without disassembling the storage bag.

[0061] In some embodiments, when executing step S4, the liquid storage bag is placed on the mixing component, and the mixing component squeezes the bottom of the storage bag by 1-2 cm at a frequency of 60-70 times / min to mix the cell suspension in the liquid storage bag evenly and prevent the cells in the cell suspension from settling at the bottom of the liquid storage bag.

[0062] In some embodiments, when executing step S4, the liquid storage bag is placed on a cooling component, and the cooling component cools the liquid storage bag to 2-8°C.

[0063] In some embodiments, when the peristaltic pump rotates forward in step S2 to pump the pre-filling liquid with a volume of V1 into the filling bag, the forward rotation speed of the peristaltic pump is 20-30 rpm.

[0064] In some embodiments, when the peristaltic pump is reversed in step S3 to pump the pre-filling liquid with a volume of V2 in the sub-packaging bag into the pre-filling bag, the reverse rotation speed of the peristaltic pump is 20-30 rpm.

[0065] In some embodiments, when the peristaltic pump rotates forward to pump the cell suspension into the filling bag in step S4, the forward rotation speed of the peristaltic pump is 20-30 rpm.

[0066] In some embodiments, the process of executing steps S2-S4 further includes: a liquid flow meter provided on the main pipeline counts the volume of the medium flowing into the sub-packaging bag, so as to help control the volume of the medium entering the sub-packaging bag.

[0067] In some embodiments, a bubble monitor is provided on the main pipeline, and the process of executing step S4 also includes: the bubble monitor provided on the main pipeline monitors whether there is gas passing through the main pipeline.

[0068] Example 1

[0069] The cell packaging method in this embodiment 1 comprises the following steps:

[0070] S1. Pre-filling pipeline connection: 10 sub-packaging bags with a nominal capacity of 20 mL are connected to a pre-filling bag containing purified water through a main pipeline, and the connection point between the main pipeline and the sub-packaging bag is located at the top of the sub-packaging bag; wherein the volume of purified water in the pre-filling bag is 200 mL;

[0071] S2. Pre-filling with priming liquid: Open the corresponding pipeline solenoid valves on 10 sub-packaging bags in turn, and when the pipeline solenoid valves are opened but not closed, the peristaltic pump pumps 10 mL of purified water into the sub-packaging bags at a forward speed of 25 rpm until all 10 sub-packaging bags are filled with purified water;

[0072] S3, exhaust of sub-packaging bags: open the corresponding pipeline solenoid valves on 10 sub-packaging bags in turn, and when the pipeline solenoid valves are opened but not closed, the peristaltic pump pumps 10 mL of the pre-filling liquid in the sub-packaging bag into the pre-filling bag at a reverse speed of 25 rpm until 20 mL of the pre-filling liquid enters the pre-filling bag in all 10 sub-packaging bags;

[0073] S4. Cell suspension subpackaging: After disconnecting the main pipeline from the pre-filled bag, connect the main pipeline to a storage bag containing 230 mL of MSC cell suspension at 3°C, and use the mixing component to squeeze the bottom of the storage bag by 2 cm at a frequency of 65 times / min. Open the corresponding pipeline solenoid valves on 10 subpackaging bags in turn, and when the pipeline solenoid valves are opened but not closed, use the peristaltic pump to pump 20 mL of cell suspension in the storage bag into the subpackaging bag at a forward speed of 25 rpm until 20 mL of cell suspension is pumped into all 10 subpackaging bags.

[0074] It was measured that after performing the above steps on 10 packaging bags, it took a total of 5.1 minutes (excluding pipeline installation time), and the volume of residual liquid in the pipeline was 14 mL.

[0075] Performance Testing

[0076] The presence of bubbles in the packaging bag in Example 1 was observed visually, and the results are shown in Table 1.

[0077] The solutions in the 10 sub-packaging bags in Example 1 were taken out and transferred into a measuring cylinder, the volume in each sub-packaging bag was measured, and the packing accuracy was calculated. The results are shown in Table 1.

[0078] Take 20 μL of the cell suspension in the storage bag, mix it with 20 μL of 0.4% trypan blue in a ratio of 1:1, and calculate the cell number and cell viability in the cell suspension; the cell amount and cell viability of the cell suspension in the 10 sub-packaging bags in Example 1 were calculated in the same manner, and the results are shown in Table 2, wherein the target cell number is the total number of cells in the storage bag multiplied by 2 / 23.

[0079] Table 1 Whether there are bubbles in the packaging bag, the filling volume and the packaging accuracy in Example 1

[0080]

[0081] Table 2 Cell number and cell viability in the packaging bag in Example 1

[0082]

[0083] Although the embodiments of the present invention are described in detail above, it is obvious to those skilled in the art that various modifications and variations can be made to these embodiments. However, it should be understood that such modifications and variations are within the scope and spirit of the present invention as described in the claims. Moreover, the present invention described herein may have other embodiments and may be implemented or realized in a variety of ways.

Claims

1. A cell packaging method, characterized in that: The following steps are involved: The sub-packaging bag with a capacity of V is connected to the pre-filling bag storing the pre-filling liquid through a main pipeline, and the connection point between the main pipeline and the sub-packaging bag is located at the top of the sub-packaging bag; Open the corresponding pipeline solenoid valve on the sub-packaging bag, and after the peristaltic pump rotates forward to pump V1 volume of pre-filling liquid into the sub-packaging bag, the pipeline solenoid valve is closed; where V1: V = 1 / 5-3 / 5; The corresponding pipeline solenoid valve on the sub-packaging bag is opened, and the peristaltic pump is reversed to pump V2 volume of the pre-filling liquid in the sub-packaging bag into the pre-filling bag, and then the pipeline solenoid valve is closed; Among them, V2: V1 = 8 / 10-9 / 10; The main pipeline is disconnected from the pre-filled bag and connected to a storage bag storing a cell suspension, the corresponding pipeline solenoid valve on the sub-packaging bag is opened, the peristaltic pump rotates forward to pump the cell suspension into the sub-packaging bag, and then the pipeline solenoid valve is closed; wherein the cell suspension is composed of cells and a resuspension medium, and the resuspension medium is of the same type as the pre-filled liquid.

2. The cell packaging method according to claim 1, characterized in that: The step of connecting the sub-packaging bag with a capacity of V to the pre-filling bag storing the pre-filling liquid through the main pipeline includes: N sub-packaging bags with a capacity of V are connected to a pre-filling bag storing pre-filling liquid through a main pipeline, wherein N is a positive integer greater than or equal to 2.

3. The cell packaging method according to claim 2, characterized in that: The volume of the pre-filled liquid in the pre-filled bag is greater than N times V.

4. The cell packaging method according to claim 2, characterized in that: The step of opening the corresponding pipeline solenoid valve on the sub-packaging bag, and then the peristaltic pump rotates forward to pump V1 volume of pre-filling liquid into the sub-packaging bag, and then closing the pipeline solenoid valve includes: The corresponding pipeline solenoid valves on the N sub-packaging bags are opened and closed in sequence. When the pipeline solenoid valve is opened but not closed yet, the peristaltic pump rotates forward to pump V1 volume of pre-filling liquid into the sub-packaging bag until all N sub-packaging bags are filled with pre-filling liquid.

5. The cell packaging method according to claim 4, characterized in that: The step of opening the corresponding pipeline solenoid valve on the sub-packaging bag, reversing the peristaltic pump to pump V2 volume of the priming liquid in the sub-packaging bag into the priming bag, and then closing the pipeline solenoid valve includes: The corresponding pipeline solenoid valves on the N sub-packaging bags are opened and closed in sequence. When the pipeline solenoid valve is opened but not closed yet, the peristaltic pump reverses to pump V2 volume of pre-filling liquid in the sub-packaging bag into the pre-filling bag until V2 volume of pre-filling liquid in each of the N sub-packaging bags is pumped into the pre-filling bag.

6. The cell packaging method according to claim 5, characterized in that: The process of opening the corresponding pipeline solenoid valve on the sub-packaging bag, rotating the peristaltic pump forward to pump the cell suspension into the sub-packaging bag, and closing the pipeline solenoid valve includes: The corresponding pipeline solenoid valves on the N sub-packaging bags are opened and closed in sequence. When the pipeline solenoid valve is opened but not closed, the peristaltic pump rotates forward to pump the cell suspension into the sub-packaging bag until the cell suspension is stored in all the N sub-packaging bags.

7. The cell packaging method according to claim 1, characterized in that: The step of disconnecting the main pipeline from the pre-filled bag and connecting it to the storage bag storing the cell suspension, opening the corresponding pipeline solenoid valve on the sub-packaging bag, and closing the pipeline solenoid valve after the peristaltic pump rotates forward to pump the cell suspension into the sub-packaging bag also includes: The storage bag is placed on the mixing component, and the mixing component squeezes the bottom of the storage bag by 1-2 cm at a frequency of 60-70 times / min.

8. The cell packaging method according to claim 1, characterized in that: When the peristaltic pump rotates forward to pump V1 volume of pre-filling liquid into the dispensing bag, the forward speed of the peristaltic pump is 20-30rpm; when the peristaltic pump rotates reversely to pump V2 volume of pre-filling liquid in the dispensing bag into the pre-filling bag, the reverse speed of the peristaltic pump is 20-30rpm; when the peristaltic pump rotates forward to pump the cell suspension into the dispensing bag, the forward speed of the peristaltic pump is 20-30rpm.

9. The cell packaging method according to claim 1, characterized in that: The step of disconnecting the main pipeline from the pre-filled bag and connecting it to the storage bag storing the cell suspension, opening the corresponding pipeline solenoid valve on the sub-packaging bag, and closing the pipeline solenoid valve after the peristaltic pump rotates forward to pump the cell suspension into the sub-packaging bag also includes: The liquid flow meter installed on the main pipeline counts the volume of the cell suspension flowing into the sub-packaging bag.

10. The cell packaging method according to claim 1, characterized in that: The step of disconnecting the main pipeline from the pre-filled bag and connecting it to the storage bag storing the cell suspension, opening the corresponding pipeline solenoid valve on the sub-packaging bag, and closing the pipeline solenoid valve after the peristaltic pump rotates forward to pump the cell suspension into the sub-packaging bag also includes: The bubble monitor installed on the main pipeline monitors whether there is gas passing through the main pipeline.

Images