Cell partitioning method

Abstract

The application provides a cell sub-packaging method, and relates to the technical field of cell sub-packaging. The sub-packaging method comprises the following steps: connecting a sub-packaging bag and a pre-charging bag storing pre-charging liquid through a total pipeline; opening a corresponding pipeline electromagnetic valve on the sub-packaging bag, and then reversely rotating a peristaltic pump to pump the pre-charging liquid in the sub-packaging bag into the pre-charging bag; opening a corresponding pipeline electromagnetic valve on the sub-packaging bag, and then reversely rotating a peristaltic pump to pump the pre-charging liquid in the sub-packaging bag into the pre-charging bag; connecting the total pipeline to a storage bag storing cell suspension after disconnecting the total pipeline from the pre-charging bag, opening a corresponding pipeline electromagnetic valve on the sub-packaging bag, and then reversely rotating a peristaltic pump to pump the cell suspension in the sub-packaging bag into the pre-charging bag. The method can be applied to the sub-packaging operation of cell preparations, can not only remove air in the sub-packaging bag, but also reduce labor, has low pollution risk, and has high automation degree.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of cell sub-packaging, in particular to a cell sub-packaging method. BACKGROUND

[0002] The production of clinical cell preparations is a complex process that employs a variety of technical means, designs multiple steps across different geographical locations, and any error in production control, storage or logistics transportation will cause cell damage, directly affecting the final patient treatment efficiency. Among them, the sub-packaging of cell preparations is a key step and starting step of cell production, which has an important impact on the quality of cell products.

[0003] However, the current sub-packaging of cell preparations is mostly manually operated, which is difficult to remove the air and other gases in the sub-packaging bag, and thus part of the air remaining in the sub-packaging bag after sub-packaging of the cell preparation will cause irreversible damage to the cells. Therefore, it is necessary to propose a solution to improve the above problems. SUMMARY

[0004] The purpose of the present application is to provide a cell sub-packaging method which can be applied to the sub-packaging operation of cell preparations, not only capable of removing the air in the sub-packaging bag, but also capable of reducing labor, low pollution risk and high automation degree.

[0005] The cell sub-packaging method provided by the present application adopts the following technical solution, comprising the following steps:

[0006] The sub-packaging bag with a capacity of V is connected in communication with the pre-charging bag storing the pre-charging liquid through a total pipeline, and the communication part of the total pipeline with the sub-packaging bag is located at the top of the sub-packaging bag;

[0007] The corresponding pipeline electromagnetic valve on the sub-packaging bag is opened, the peristaltic pump is rotated to pump V1 volume of pre-charging liquid into the sub-packaging bag, and then the pipeline electromagnetic valve is closed; wherein V1:V=1 / 5-3 / 5;

[0008] The corresponding pipeline electromagnetic valve on the sub-packaging bag is opened, the peristaltic pump is reversed to pump V2 volume of pre-charging liquid in the sub-packaging bag into the pre-charging bag, and then the pipeline electromagnetic valve is closed; wherein V2:V1=8 / 10-9 / 10;

[0009] After the total pipeline is disconnected from the pre-charging bag and connected to the storage bag storing the cell suspension, the corresponding pipeline electromagnetic valve on the sub-packaging bag is opened, the peristaltic pump is rotated to pump the cell suspension into the sub-packaging bag, and then the pipeline electromagnetic valve is closed; wherein the cell suspension is composed of cells and resuspension medium, and the resuspension medium and the pre-charging liquid are of the same type.

[0010] The cell sub-packaging method has the beneficial effects that: when the cell suspension is sub-packaged, the pre-priming bag storing the pre-priming liquid is connected with the sub-packaging bag in advance, the peristaltic pump pumps the pre-priming liquid into the sub-packaging bag, and the ratio of the volume V1 of the pre-priming liquid pumped into the sub-packaging bag to the capacity V of the sub-packaging bag is 1 / 5-3 / 5, since the communication position of the sub-packaging bag and the total pipeline is located at the top of the sub-packaging bag, the pre-priming liquid is located at the bottom position of the sub-packaging bag; when the peristaltic pump reverses to pump the pre-priming liquid in the sub-packaging bag into the pre-priming bag, since the top of the sub-packaging bag is gas, the peristaltic pump firstly pumps the gas in the sub-packaging bag into the pre-priming bag, when the peristaltic pump pumps the pre-priming liquid with the volume V2 in the sub-packaging bag into the pre-priming bag, the gas in the sub-packaging bag can be completely pumped into the pre-priming bag, so that only the pre-priming liquid remains in the sub-packaging bag, and then the peristaltic pump is forwardly rotated to pump the cell suspension into the sub-packaging bag, so that the cell suspension is not in contact with the gas such as air in the sub-packaging bag, and the resuspension medium in the cell suspension is the same as the pre-priming liquid, that is, the pre-priming liquid remaining in the sub-packaging bag does not affect the cells in the cell suspension.

[0011] Optionally, in the step of connecting the sub-packaging bag with the capacity V and the pre-priming bag storing the pre-priming liquid through the total pipeline, the step includes: connecting N sub-packaging bags with the capacity V and the pre-priming bag storing the pre-priming liquid through the total pipeline, wherein N is a positive integer greater than or equal to 2. After connecting the N sub-packaging bags with the total pipeline, the N sub-packaging bags can be pre-charged and degassed, and after degassing, the N sub-packaging bags can be sub-packaged with the cell suspension, so that the sub-packaging efficiency is improved.

[0012] Optionally, the volume of the pre-priming liquid in the pre-priming bag is greater than N times V, so that the pre-priming liquid is not insufficient when pumped into the sub-packaging bag.

[0013] Optionally, in the step of opening the corresponding pipeline electromagnetic valve of the sub-packaging bag, the peristaltic pump is forwardly rotated to pump the pre-priming liquid with the volume V1 into the sub-packaging bag, and then the pipeline electromagnetic valve is closed, the step includes: the corresponding pipeline electromagnetic valves of the N sub-packaging bags are opened and closed in sequence, and when the pipeline electromagnetic valve is opened but not closed, the peristaltic pump is forwardly rotated to pump the pre-priming liquid with the volume V1 into the sub-packaging bag, until the N sub-packaging bags all have the pre-priming liquid. The pipeline electromagnetic valves of the N sub-packaging bags are opened and closed in sequence, so that the peristaltic pump can pump the pre-priming liquid into each sub-packaging bag when working.

[0014] Optionally, the step of opening the corresponding pipeline solenoid valve on the sub-packaging bag, reversing the peristaltic pump to pump V2 volume of pre-filling liquid from the sub-packaging bag into the pre-filling bag, and then closing the pipeline solenoid valve includes: opening and closing the corresponding pipeline solenoid valves on N sub-packaging bags in sequence, and when the pipeline solenoid valves are opened but not closed yet, reversing the peristaltic pump to pump V2 volume of pre-filling liquid from the sub-packaging bag into the pre-filling bag, until V2 volume of pre-filling liquid is pumped into the pre-filling bag from each of the N sub-packaging bags. The peristaltic pump reverses to extract 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 pre-filling liquid in each of the N sub-packaging bags can be extracted into the pre-filling 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 cell suspension is stored in all N sub-packaging bags.

[0016] Optionally, the step of disconnecting the main pipeline from the pre-filled bag and connecting it to a storage bag storing the cell suspension, 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 also includes: placing the storage bag on a mixing component, and squeezing 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 evenly mix the cell suspension in the storage bag and prevent the cells in the cell suspension from settling, thereby ensuring that the cells in the cell suspension pumped out by the peristaltic pump are evenly distributed.

[0017] Optionally, when the peristaltic pump is rotated forward to pump V1 volume of pre-filling liquid into the sub-filling bag, the forward rotation speed of the peristaltic pump is 20-30 rpm; when the peristaltic pump is rotated reversely to pump V2 volume of pre-filling liquid in the sub-filling bag into the pre-filling bag, the reverse rotation speed of the peristaltic pump is 20-30 rpm; when the peristaltic pump is rotated forward to pump the cell suspension into the sub-filling bag, the forward rotation speed of the peristaltic pump is 20-30 rpm.

[0018] Optionally, the steps of disconnecting the main pipeline from the pre-filled bag and then connecting it to a storage bag containing the cell suspension, 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 may further include: using a liquid flow meter provided on the main pipeline to measure the volume of the cell suspension flowing into the sub-packaging bag. This allows for controlling the volume of the cell suspension pumped into the sub-packaging bag, thereby facilitating consistent volume control of the cell suspension within the sub-packaging bag.

[0019] Optionally, after the main pipeline is disconnected from the pre-charging bag and connected to the storage bag storing the cell suspension, the corresponding pipeline electromagnetic valve on the storage bag is opened, the peristaltic pump is rotated forward to pump the cell suspension into the storage bag, and the pipeline electromagnetic valve is closed. In this step, the bubble monitor arranged on the main pipeline monitors whether gas passes through the main pipeline. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 is a step flow chart of the cell dispensing method in the embodiment of the present application;

[0021] Figure 2 is a specific step flow chart of step S2 in the embodiment of the present application;

[0022] Figure 3 is a specific step flow chart of step S3 in the embodiment of the present application;

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

[0024] To make the objectives, technical solutions, and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below. Obviously, the described embodiments are some but not all of the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the scope of the present application. Unless otherwise defined, the technical terms or scientific terms used herein should be understood as the common meanings thereof by those of ordinary skill in the art. The similar words such as “comprise” used herein mean that the elements or objects before the words encompass the elements or objects listed after the words and their equivalents, and do not exclude other elements or objects.

[0025] The embodiments of the present application provide a cell dispensing method.

[0026] Referring to Figure 1 , the cell dispensing method comprises the following steps:

[0027] S1, pre-charging pipeline connection: a storage bag with a capacity of V is connected in communication with a pre-charging bag storing pre-charging liquid through a main pipeline, and the communication between the main pipeline and the storage bag is located at the top of the storage bag;

[0028] S2, pre-charging liquid pre-charging: the corresponding pipeline electromagnetic valve on the storage bag is opened, the peristaltic pump is rotated forward to pump V1 volume of pre-charging liquid into the storage bag, and the pipeline electromagnetic valve is closed; wherein V1:V=1 / 5-3 / 5;

[0029] S3. Exhaust 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 priming liquid in the sub-packaging bag into the priming bag, and then the pipeline solenoid valve is closed; where V2: V1 = 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. After the peristaltic pump rotates forward to pump the cell suspension into the sub-packaging bag, the pipeline solenoid valve is closed. 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 lumen volume of the main pipeline is 40 mL, that is, a maximum of 40 mL of 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 priming bag storing priming liquid through a main pipeline, where N is a positive integer greater than or equal to 2. Connecting multiple sub-packaging bags to the main pipeline allows priming liquid to be pumped into the multiple sub-packaging bags, thereby improving overall utilization 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 priming liquid in the priming bag is greater than N multiplied by V, that is, the volume of the priming liquid in the priming bag is greater than the total capacity of N sub-packaging bags, so that the priming liquid can be distributed to at least N sub-packaging bags, and the priming operation of the N sub-packaging bags with the priming liquid is completed.

[0037] 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 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 yet, the peristaltic pump rotates forward to pump V1 volume of pre-filling liquid into the sub-packaging bags until all N sub-packaging bags are filled with pre-filling 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 N sub-packaging bags contain pre-filled liquid, and the volumes of the pre-filled liquid in the N sub-packaging bags can 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-fill bag storing the pre-fill 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. The first pipeline solenoid valve is opened, and the second, third, and fourth pipeline solenoid valves remain closed. The peristaltic pump rotates forward to pump a volume V1 of priming liquid into the first sub-packaging bag. The first pipeline solenoid valve and the peristaltic pump are then closed. Before the priming liquid is pumped into the first sub-packaging bag, the priming 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, the peristaltic pump rotates forward to pump V1 volume of priming liquid into the second filling bag, and then the second pipeline solenoid valve and the peristaltic pump are closed;

[0042] S23, 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 priming liquid of volume V1 into the third sub-packaging bag, and then the solenoid valve of the third pipeline and the peristaltic pump are closed;

[0043] S24. Open the solenoid valve of the fourth pipeline, and keep the solenoid valve of the first pipeline, the solenoid valve of the second pipeline, and the solenoid valve of the third pipeline closed. After the peristaltic pump rotates forward to pump the pre-filling liquid with a volume of V1 into the fourth filling bag, the solenoid valve of the fourth pipeline and the peristaltic pump are closed.

[0044] 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 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.

[0045] In some embodiments, when N sub-packaging bags are connected to a priming bag containing priming liquid via a main pipeline, executing step S3 includes: sequentially opening and closing the corresponding pipeline solenoid valves on the N sub-packaging bags, and when the pipeline solenoid valves are opened but not closed, reversing the peristaltic pump to pump the priming liquid in the sub-packaging bags into the priming bag, until all N sub-packaging bags have priming liquid pumped into the priming bag. The volumes of priming liquid pumped into the priming bag in each of the N sub-packaging bags may be the same or different, but at least a portion of the priming liquid in each of the N sub-packaging bags is pumped into the priming 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 from 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, reverse the peristaltic pump to pump V2 volume of priming liquid in the first sub-packaging bag into the priming bag, and then close the first pipeline solenoid valve and the peristaltic pump;

[0048] S32. Open the second pipeline solenoid valve, while the first, third, and fourth pipeline solenoid valves remain closed. The peristaltic pump reverses and pumps V2 volume of priming liquid from the second sub-bag into the priming bag. Then, the second pipeline solenoid valve and the peristaltic pump are closed.

[0049] S33. Open the solenoid valve of the third pipeline, and keep the solenoid valves of the first pipeline, the second pipeline, and the fourth pipeline closed. After the peristaltic pump reverses and pumps the priming liquid with a volume of V2 in the third sub-bag into the priming bag, the solenoid valve of the third pipeline and the peristaltic pump are closed.

[0050] S34. Open the solenoid valve of the fourth pipeline, and keep the solenoid valve of the first pipeline, the solenoid valve of the second pipeline, and the solenoid valve of the third pipeline closed. After the peristaltic pump reverses to pump the priming liquid with a volume of V2 in the fourth sub-packaging bag into the priming bag, the solenoid valve of the fourth pipeline 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 all N sub-packaging bags.

[0052] In some embodiments, when the N sub-packaging bags are connected to the priming bag storing the priming 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-fill bag storing the pre-fill 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, opening the first pipeline solenoid valve, 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 sub-packaging 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 fourth pipeline solenoid valve, keep the first pipeline solenoid valve, the second pipeline solenoid valve and the third pipeline solenoid valve closed, and after the peristaltic pump rotates forward to pump the cell suspension in the storage bag into the fourth filling bag, the fourth pipeline solenoid valve and the peristaltic pump are closed.

[0059] In some embodiments, when 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 dispensed 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 assembly, and the cooling assembly 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 solution 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 priming liquid with a volume of V2 in the sub-packaging bag into the priming 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, which is helpful for controlling 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 further 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. Prefill line connection: Connect 10 20mL aliquot bags to a prefill bag containing purified water through a main line, with the main line connecting the aliquot bags at the top of the aliquot bags. The volume of purified water in the prefill bag is 200mL.

[0071] S2. Priming of priming liquid: Open the corresponding pipeline solenoid valves on 10 sub-packaging bags in sequence. After the pipeline solenoid valves are opened but not closed, the peristaltic pump is used to pump 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 the sub-packaging bags: Open the corresponding pipeline solenoid valves on the 10 sub-packaging bags in sequence. After the pipeline solenoid valves are opened but not closed, the peristaltic pump is used to pump 10 mL of the priming solution from the sub-packaging bag into the priming bag at a reverse speed of 25 rpm until 20 mL of priming solution is added to each of the 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 squeeze the bottom of the storage bag by 2 cm at a frequency of 65 times / min with the mixing component. Open the corresponding pipeline solenoid valves on 10 sub-packaging 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 sub-packaging bags at a forward speed of 25 rpm until 20 mL of cell suspension is pumped into all 10 sub-packaging bags.

[0074] It was determined that after performing the above steps on 10 packaging bags, a total of 5.1 minutes were spent (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 bags of 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 respectively taken out and transferred into graduated cylinders. The volume in each sub-packaging bag was measured, and the packing accuracy was calculated. The results are shown in Table 1.

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

[0079] Table 1 Presence of bubbles in the packaging bag, filling volume and packaging accuracy in Example 1

[0080]

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

[0082]

[0083] While the embodiments of the present invention have been described in detail above, it will be apparent to those skilled in the art that various modifications and variations of these embodiments are possible. However, it should be understood that such modifications and variations are within the scope and spirit of the present invention as set forth in the claims. Furthermore, the invention described herein is susceptible to other embodiments and may be practiced or implemented in a variety of ways.

Claims

1. A cell packaging method, characterized in that: The following steps are involved: Connecting a sub-packaging bag with a capacity of V to a pre-filling bag storing a pre-filling liquid through a main pipeline, wherein 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; Open the corresponding pipeline solenoid valve on the sub-packaging bag, and the peristaltic pump reverses to pump V2 volume of priming liquid in the sub-packaging bag into the priming bag, and then the pipeline solenoid valve is closed; Among them, V2: V1 = 8 / 10-9 / 10; 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-filling bag. After the peristaltic pump rotates forward to pump the cell suspension into the sub-filling bag, the pipeline solenoid valve is closed. The cell suspension consists of cells and 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, wherein The step of connecting the sub-filling bag with a capacity of V to the priming bag storing the priming liquid through a main pipeline includes: N sub-packaging bags with a capacity of V are connected to a pre-filling bag storing a pre-filling liquid through a main pipeline, where 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 priming liquid in the priming bag is greater than N multiplied by V.

4. The cell packaging method according to claim 2, wherein The steps of opening the corresponding pipeline solenoid valve on the sub-packaging bag, pumping V1 volume of pre-filling liquid into the sub-packaging bag by the peristaltic pump in the forward direction, and then closing the pipeline solenoid valve include: 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-filling bag, reversing the peristaltic pump to pump V2 volume of priming liquid in the sub-filling 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 is pumped into the pre-filling bag in each of the N sub-packaging bags.

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 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 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 steps 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-filling bag, rotating the peristaltic pump forward to pump the cell suspension into the sub-filling bag, and then closing the pipeline solenoid valve also include: 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, wherein When the peristaltic pump rotates forward to pump V1 volume of pre-filling liquid into the sub-packaging bag, the forward speed of the peristaltic pump is 20-30 rpm; when the peristaltic pump rotates reversely to pump V2 volume of pre-filling liquid in the sub-packaging bag into the pre-filling bag, the reverse speed of the peristaltic pump is 20-30 rpm; when the peristaltic pump rotates forward to pump the cell suspension into the sub-packaging bag, the forward speed of the peristaltic pump is 20-30 rpm.

9. The cell packaging method according to claim 1, characterized in that: The steps 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-filling bag, rotating the peristaltic pump forward to pump the cell suspension into the sub-filling bag, and then closing the pipeline solenoid valve also include: 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 steps 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-filling bag, rotating the peristaltic pump forward to pump the cell suspension into the sub-filling bag, and then closing the pipeline solenoid valve also include: The bubble monitor installed on the main pipeline monitors whether there is gas passing through the main pipeline.

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