Fully automatic suspension cell culture bag kit

Abstract

Provided is a fully automatic suspension cell culture bag kit, comprising a primary cell storage bag, an initial cell activation bag, a liquid waste bag and a cell culture bag, wherein the bags are connected to each other by means of T-joints and a plurality of pipes. The cell culture bag kit eliminates the need for the manual liquid transfer operation of transferring cells into the bags from a bottle during a suspension cell culture process, such that the risk of bacterial contamination during the cell culture process is significantly reduced.

Description

A fully automatic suspension cell culture bag set

[0001] This application claims the benefit of priority to Chinese Patent Application Nos. 2023218446223, 2023108631538, 202310860536X, 202310864234X, 2023218491534, 2023218446257, 2023218491708 filed on July 14, 2023, and 2024211814534 filed on May 28, 2024, the disclosures of which are incorporated herein by reference as if fully written herein. Technical Field

[0002] The present invention relates to the field of cell culture, in particular to a culture bag set for culturing suspended cells, and more particularly to bag structures with different uses and related connection methods. Background Art

[0003] Traditional suspension cell culture (such as NK cells, 293, CHO, K562) is mainly carried out using a cell culture flask plus a cell culture bag. The initial cells / inoculation and the corresponding culture medium are added to the cell culture flask or cell culture bag to form a cell culture environment, and the cells are maintained in a suitable environment for growth and reproduction. At present, when the initial amount of cells is low, cell culture flasks are used for culture. When the number of cells is sufficient, they are transferred to a new culture bag for culture until the required number of cells is expanded. This culture method is mainly based on manual operation and requires multiple transfer operations of liquid and culture medium. The operation is relatively cumbersome and easily increases the risk of bacterial contamination.

[0004] During cell culture, culture medium must be added from bottles or bags to culture bags. Currently, connecting different bags using male and female Luer connectors can lead to entanglement of tubing. Therefore, a more secure and reliable connection method is needed to improve operational efficiency and safety.

[0005] Existing pipes between different bags need to rely on threads for connection. After being connected for a long time, due to the certain rotational force of the pipes, the threaded connections may fall off or leak, which can easily lead to contamination and affect the actual cell culture results.

[0006] Summary of the Invention

[0007] In view of the technical problems raised in the background technology, the present invention provides a fully automatic suspension cell activation culture kit to solve at least one of the technical problems mentioned in the background technology.

[0008] In order to achieve the above purpose, the present invention provides the following technical solutions:

[0009] The present invention relates to a fully automatic suspension cell activation culture kit, wherein the cell culture bag kit includes:

[0010] A primary cell storage bag is used to store primary cells, and the primary cell storage bag includes a primary cell storage bag body, an injection port, and a transfer port;

[0011] An initial cell activation bag is used to activate initial suspended cells, and the initial cell activation bag includes an initial cell activation bag body, a coating liquid injection port, a culture medium and cell liquid inlet, and a liquid transfer port;

[0012] A waste liquid bag, used to collect waste liquid in the initial cell activation bag, the waste liquid bag comprising a waste liquid bag body and a liquid inlet;

[0013] A cell culture bag is used for large-volume cell culture, comprising a cell culture bag body, a liquid inlet, and a sampling port;

[0014] Among them, the transfer outlet of the primary cell storage bag is connected to the culture medium and cell inlet of the initial cell activation bag through pipelines, and the liquid transfer outlet of the initial cell activation bag is connected to the waste liquid bag inlet and the cell culture bag inlet through pipelines respectively.

[0015] As a further preferred technical solution, at least one of the injection port of the primary cell storage bag and the coating solution injection port of the initial cell activation bag adopts a heparin sodium cap.

[0016] As a further preferred technical solution, the cell culture bag further comprises a cell density detection port, which is made of a light-transmitting material and can detect the cell density through the absorbance of the liquid in the culture bag.

[0017] As a further preferred technical solution, it also includes a four-way connector, three connectors of which are respectively connected to three culture medium inlet pipelines, and the fourth connector of the four-way connector is connected to the culture medium and cell inlet of the initial cell activation bag through a pipeline.

[0018] As a further preferred technical solution, it also includes a three-way connector A, which is used to connect the pipeline connected to the transfer outlet of the primary cell storage bag, the pipeline connected to the fourth connector of the four-way connector, and the culture medium and cell inlet pipeline of the initial cell activation bag.

[0019] As a further preferred technical solution, it further includes a three-way connector B for connecting the liquid transfer outlet pipeline of the initial cell activation bag, the liquid inlet pipeline of the waste liquid bag and the liquid inlet pipeline of the cell culture bag.

[0020] As a further preferred technical solution, at least one of the primary cell storage bag transfer outlet, the culture medium and cell inlet and liquid transfer outlet of the initial cell activation bag, and the liquid inlet of the cell culture bag is connected to the pipeline using a Luer connector.

[0021] As a further preferred technical solution, the pipeline adopts a silicone tube.

[0022] As a further preferred technical solution, at least one of the primary cell storage bag, initial cell activation bag, waste liquid bag and cell culture bag is made of FEP waterproof and breathable film material.

[0023] As a further preferred technical solution, the volume capacity of the cell culture bag is 2 L or more.

[0024] As a further preferred technical solution, the cell density detection port is made of FEP material and includes an upper light-transmitting hole and a lower light-transmitting hole that are opposed to each other, and the upper light-transmitting hole and the lower light-transmitting hole are blind holes.

[0025] As a further preferred technical solution, each corner of at least one of the primary cell storage bag, the initial cell activation bag and the cell culture bag is rounded.

[0026] As a further preferred technical solution, the initial cell activation bag is in the shape of a convex character with a narrow upper end and a wide lower end. The corners of the convex inner cavity of the convex initial cell activation bag are rounded, and the coating liquid injection port, culture medium and cell liquid inlet, and liquid transfer port are arranged at the upper end of the convex character.

[0027] As a further preferred technical solution, a quick-connect connector is also included, including a male Luer connector and a female Luer connector that cooperates with the male Luer connector, wherein:

[0028] The male Luer connector includes a first male fitting portion, a first clamping plate, a first gripping portion, a first pipe connector, and a first channel running through the first male fitting portion, the first clamping plate, the first gripping portion, and the first pipe connector, wherein the first pipe connector is used to connect to a pipeline;

[0029] The female Luer connector includes a first female fitting portion, a first clamping portion, and a second pipe connector. The first female fitting portion is provided with a first cavity into which the first male fitting portion is inserted. The second pipe connector is provided with a second channel that is in communication with the first cavity. The second pipe connector is used to connect to a pipeline.

[0030] The first clamping plate and the first clamping portion can be clamped and connected to fix the male Luer connector and the female Luer connector and connect the first channel and the second channel;

[0031] The quick-connector is arranged on the pipeline between the four-way connector and the three-way connector A, and / or the pipeline between the three-way connector B and the liquid inlet of the cell culture bag, and / or the end of the culture medium liquid inlet pipeline.

[0032] As a further preferred technical solution, the first clamping portion is at least two first clamping blocks with L-shaped cross-sections provided on the end surface of the first female matching portion, and the first clamping plate is provided with at least two first notches corresponding to the positions of the first clamping blocks. The first clamping blocks can pass over the first notches and be clamped to the first clamping plate after being rotated a certain angle to clamp the male Luer connector and the female Luer connector together.

[0033] As a further preferred technical solution, at least one first annular groove is provided on the outer periphery of the first male mating portion, a first sealing ring is provided in the first annular groove, and the first sealing ring is press-fitted with the inner wall of the first cavity of the first female mating portion;

[0034] And / or, a first sealing ring is provided on the end surface of the first male fitting portion facing the female Luer connector, and the first sealing ring can be compressed between the bottom wall of the first chamber of the female Luer connector and the end surface of the first male fitting portion.

[0035] As a further preferred technical solution, it also includes a male Luer plug that is connected to the female Luer connector. The male Luer plug includes a second male matching portion, a second clamping plate, and a second gripping portion. The second clamping plate is provided with at least two second notches corresponding to the position of the first clamping block. The first clamping block can pass over the second notches and be clamped to the second clamping plate after being rotated a certain angle to clamp the male Luer plug and the female Luer connector together.

[0036] As a further preferred technical solution, at least one second annular groove is provided on the outer circumference of the second male mating portion, a second sealing ring is provided in the second annular groove, and the second sealing ring is press-fitted with the inner wall of the first cavity of the first female mating portion;

[0037] And / or, a second sealing ring is provided on the end surface of the second male fitting portion facing the female Luer connector, and the second sealing ring can be compressed between the bottom wall of the first chamber of the female Luer connector and the end surface of the second male fitting portion.

[0038] As a further preferred technical solution, a female Luer plug is further included to be connected to the male Luer connector. The female Luer plug includes a second female fitting portion and a second clamping portion. The second clamping portion is at least two second clamping blocks with an L-shaped cross-section provided on the end surface of the second female fitting portion. The second clamping blocks can pass over the first notch and be clamped to the first clamping plate after being rotated a certain angle to clamp the female Luer plug and the male Luer connector together.

[0039] As a further preferred technical solution, the first clamping plate between adjacent first notches has a thickened wedge-shaped inclined surface or a trapezoidal inclined surface on one side facing the first gripping portion;

[0040] And / or, a surface of the second clamping plate between adjacent second notches facing the second gripping portion has a thickened wedge-shaped inclined surface or a trapezoidal inclined surface.

[0041] The patent of this invention solves the shortcoming of existing suspension cell culture that requires multiple manual liquid transfers. It provides a packaged system for fully automatic suspension cell activation culture, which ensures a sterile environment for cell culture and can cooperate with the equipment to observe cell density without sampling, overcoming the risk of cross-contamination of cells in manual operation projects.

[0042] Furthermore, using the quick-connect connector of the present invention, when connecting two sections of pipelines, the male Luer connector and the female Luer connector are quickly connected through the first clamping plate and the first clamping portion to achieve pipeline connectivity. Only a small rotation angle is required to achieve pipeline connection, preventing the pipelines from becoming entangled. Furthermore, the rotating clamping structure makes the connection faster and more secure, thereby improving the efficiency of liquid transfer. This quick-connect method can achieve a stable and tight connection between two different bag bodies. Furthermore, when liquid transfer is not required, the male Luer connector and the female Luer connector are disconnected, and then the male Luer plug is quickly connected to the female Luer connector to block the pipeline on the female Luer connector side, preventing liquid leakage in the pipeline on the female Luer connector side. The female Luer plug is quickly connected to the male Luer connector to block the pipeline on the male Luer connector side, preventing liquid leakage in the pipeline on the male Luer connector side. The structure is simpler and easier to operate, greatly improving the effectiveness of pipeline connection while reducing the risk of contamination.

[0043] BRIEF DESCRIPTION OF THE DRAWINGS

[0044] FIG1 is a schematic structural diagram of a fully automatic suspension cell culture bag set according to an embodiment of the present invention;

[0045] FIG2 is a schematic structural diagram of one side of a cell density detection port in the present invention;

[0046] FIG3 is a schematic structural diagram of the other side of the cell density detection port in the present invention;

[0047] FIG4 is a top view of a cell density detection port in the present invention;

[0048] FIG5 is a cross-sectional view taken along AA in FIG4 ;

[0049] FIG6 is a schematic structural diagram of a fully automatic suspension cell culture bag set according to another embodiment of the present invention;

[0050] FIG7 is a schematic structural diagram of a male Luer connector of a quick-connect connector used in the present invention;

[0051] FIG8 is a schematic structural diagram of a female Luer connector of a quick-connect connector used in the present invention;

[0052] FIG9 is a schematic structural diagram of a male Luer connector and a female Luer connector used in the present invention after being connected;

[0053] FIG10 is a schematic structural diagram of a male Luer plug of a quick-connect connector used in the present invention;

[0054] FIG11 is a schematic structural diagram of a quick-connect female Luer connector and a male Luer plug used in the present invention after being connected;

[0055] FIG12 is a schematic structural diagram of a female Luer plug of a quick-connect connector used in the present invention;

[0056] FIG13 is a schematic structural diagram of a quick-connect male Luer connector and a female Luer plug used in the present invention after being connected.

[0057] In the figure: 1-primary cell storage bag, 101-primary cell storage bag body, 102-injection port, 103-transfer port, 2-initial cell activation bag, 201-initial cell activation bag body, 202-coating solution injection port, 203-culture medium and cell liquid inlet, 204-liquid transfer port, 3-waste liquid bag, 301-waste liquid bag body, 302-liquid inlet, 4-cell culture bag, 401-cell culture bag body, 402-liquid inlet, 403-sampling port, 404-cell density detection port, 4041-connecting part, 40411-through hole, 4042-detection part, 40421-cavity, 40431-upper light-transmitting hole, 40432-lower light-transmitting hole, 501-507-pipeline, 601-tee connector A, 602-tee connector B, 701-four-way connector, 8, 8'-quick plug Connector, 81-male Luer connector, 811-first male fitting part, 8111-first annular groove, 812-first clamping plate, 8121-first notch, 813-first gripping part, 814-first pipe connector, 815-first channel, 82-female Luer connector, 821-first female fitting part, 8211-end face, 822-first clamping part, 8221-first clamping block, 823-second pipe connector, 824-first chamber, 825-second channel, 83-male Luer plug, 831-second male fitting part, 8311-second annular groove, 832-second clamping plate, 8321-second notch, 833-second gripping part, 84-female Luer plug, 841-second female fitting part, 8411-end face, 842-second clamping part, 8421-second clamping block, 843-second chamber. Best Mode for Carrying Out the Invention

[0058] The features and exemplary embodiments of various aspects of the present invention will be described in detail below. In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention will be further described in detail with reference to the accompanying drawings and examples. It should be understood that the specific embodiments described herein are only configured to explain the present invention and are not configured to limit the present invention. For those skilled in the art, the present invention can be implemented without the need for some of these specific details. The following description of the embodiments is merely intended to provide a better understanding of the present invention by illustrating examples of the present invention.

[0059] It should be noted that, in this document, relational terms such as first and second, etc., are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "comprises," "comprising," or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article, or device comprising a series of elements includes not only those elements, but also other elements not explicitly listed, or elements inherent to such process, method, article, or device. In the absence of further limitations, the elements defined by the phrase "comprising..." do not exclude the presence of other identical elements in the process, method, article, or device comprising the elements.

[0060] Example 1

[0061] Figure 1 shows a fully automatic suspension cell culture bag set according to the present invention, comprising a primary cell storage bag 1, an initial cell activation bag 2, a waste fluid bag 3, and a cell culture bag 4. Preferably, the bag bodies of the primary cell storage bag 1, initial cell activation bag 2, waste fluid bag 3, and cell culture bag 4 are made of FEP waterproof, breathable membrane. FEP waterproof, breathable membrane is chemically stable, non-cytotoxic, and non-tissue toxic, promoting cell proliferation. Furthermore, its transparent material facilitates microscopic observation.

[0062] The primary cell storage bag 1 is used to store primary cells and preferably has a volume capacity of 20 ml. The primary cell storage bag 1 includes a primary cell storage bag body 101, an injection port 102, and a transfer port 103. The initial cell activation bag 2 is used to activate initial suspended cells and includes an initial cell activation bag body 201, a coating solution injection port 202, a culture medium and cell inlet 203, and a liquid transfer port 204. The waste liquid bag 3 is used to collect waste liquid from the initial cell activation bag and includes a waste liquid bag body 301 and a liquid inlet 302. The cell culture bag 4 is used for large-volume cell culture and preferably has a volume capacity of 2 L or more. It includes a cell culture bag body 401, a liquid inlet 402, and a sampling port 403.

[0063] The primary cell storage bag 1, initial cell activation bag 2, waste liquid bag 3 and cell culture bag 4 are connected via a three-way connector A 601, a three-way connector B 602 and a plurality of pipes 501-507. The pipes 501-507 are preferably silicone tubes.

[0064] Specifically, the transfer outlet 103 of the primary cell storage bag 1 is connected to the culture medium and cell liquid inlet 203 of the initial cell activation bag 2 through pipelines 501 and 502, and the liquid transfer outlet 204 of the initial cell activation bag 2 is connected to the liquid inlet 302 of the waste liquid bag 3 and the liquid inlet 402 of the cell culture bag through pipelines 503, 504, and 505, respectively.

[0065] Pipelines 501 and 502 are connected via a tee connector A 601 , and pipelines 503 , 504 , and 505 are connected via a tee connector B 602 .

[0066] Furthermore, the injection port 102 of the primary cell storage bag 1 and the coating fluid injection port 202 of the initial cell activation bag 2 utilize sodium heparin caps. The injection port allows for manual addition of primary cells into the bag, minimizing contact between the bag interior and the external environment and reducing the risk of contamination. The injection port allows for manual addition of coating fluid, allowing for direct addition of fluid into the bag, preventing loss of coating fluid in the piping. This facilitates more effective coating of the bag material and enhances initial cell activation.

[0067] Furthermore, the cell culture bag 4 further includes a cell density detection port 404 . The cell density detection port 404 is made of a light-transmitting material and can detect the cell density through the absorbance of the liquid in the culture bag.

[0068] Specifically, as shown in FIG2 , the cell density detection port 404 includes a connecting portion 4041 and a detecting portion 4042 , wherein the connecting portion 4041 is used for sealing connection with the cell culture bag 4 . More preferably, the cell density detection port 404 is made of FEP material.

[0069] Preferably, the connecting portion 4041 is diamond-shaped, and a through hole 40411 is provided in the middle thereof to communicate with the cavity 40421 of the detecting portion 4042 .

[0070] Further preferably, the detection portion 4042 includes a bottom wall, an upper wall, a lower wall, and an arcuate sidewall connecting the upper and lower walls. The upper and lower walls are preferably flat. Preferably, the upper and lower walls are provided with an upper light-transmitting hole 40431 and a lower light-transmitting hole 40432, which are opposed in the vertical direction. Both the upper light-transmitting hole 40431 and the lower light-transmitting hole 40432 are blind holes, that is, recessed grooves in the upper and lower wall surfaces that do not pass through the cavity 40421. Further preferably, the aperture of the upper light-transmitting hole 40431 and the lower light-transmitting hole 40432 is 2-4 mm, more preferably 3 mm, and the thickness at the light-transmitting hole is 0.1-0.3 mm, more preferably 0.2 mm.

[0071] The cell density detection port 404 made of FEP material of the present invention has the advantages of good light transmittance and repeatability compared to ordinary plastic materials. For the same culture sample, the light transmittance intensity at the cell density detection port was tested multiple times. The test results of cell density detection ports made of different materials are detailed in the comparison table below.

[0072] For the same culture sample, at different culture days, the cell density was measured non-contact using the absorbance of the culture medium at the cell density detection port. There was no substantial difference in the results compared to the traditional method of removing the culture medium and then measuring the cell density using a flow cytometer. See the comparison table below for details.

[0073] Furthermore, the cell culture bag set further includes a four-way connector 701, three connectors of which are respectively connected to three culture medium inlet pipes 506, and the fourth connector of the four-way connector is connected to the culture medium and cell inlet 203 of the initial cell activation bag 2 through pipes 507, 502, and three-way connector A 601.

[0074] Furthermore, the three-way connector A 601 is connected to the pipeline 501 connected to the transfer outlet 103 of the primary cell storage bag 1 , the pipeline 507 connected to the fourth connector of the four-way connector 701 , and the culture medium and cell inlet pipeline 502 of the initial cell activation bag 2 .

[0075] The three-way connector B 602 is connected to the liquid outlet pipe 503 of the initial cell activation bag 2 , the waste liquid bag liquid inlet pipe 504 , and the cell culture bag liquid inlet pipe 505 .

[0076] Furthermore, the primary cell storage bag transfer outlet 103, the culture medium and cell liquid inlet 203 and liquid transfer outlet 204 of the initial cell activation bag, and the liquid inlet 402 of the cell culture bag are connected to the pipeline using Luer connectors.

[0077] Example 2

[0078] 6 shows another fully automatic suspension cell culture bag set of the present invention. In this embodiment, the same components as those in the first embodiment use the same names and labels, and the repeated components are not described again. The following focuses on the details that are different from the first embodiment.

[0079] Specifically, the initial cell activation bag 2 adopts a special shape relative to the first embodiment, so that the edge of the inner cavity of the initial cell activation bag 2 is rounded to avoid dead corners. Further preferably, the initial cell activation bag 2 is specifically convex (or gourd-shaped), see Figure 6, that is, the upper end is narrow and the lower end is wide. Preferably, a convex-shaped initial cell activation bag body 201 is processed on a rectangular or square bag body by heat welding, and the middle of the body 201 is a cavity for accommodating initial cells; more preferably, the corners of the inner cavity of the convex initial cell activation bag are rounded, and the two inner convex corners can be right angles or rounded. The convex upper end of the initial cell activation bag 2 is provided with a coating liquid injection port 202, a culture medium and cell liquid inlet 203, and a liquid transfer port 204.

[0080] In this embodiment, the convex-shaped initial cell activation bag 2 has better effects than the rectangular bag body in Example 1: 1. When welding the interfaces of the coating liquid injection port 202, the culture medium and cell liquid inlet 203, and the liquid transfer port 204 of the rectangular bag body, the overall bag is severely wrinkled, and the wrinkles will be more serious after sterilization; and during coating and activation, a necessary condition is to keep the interface with the cell inoculation flat, otherwise the wrinkles of the bag will cause cell aggregation, affecting the cell and the cytokine inoculation at the bottom and affecting the degree of activation. Therefore, in order to ensure the flatness of the bottom of the bag, it is designed to be a convex-shaped bag with a narrow upper end and a wide lower end. Even after welding the interfaces at the upper end, the lower end will not be wrinkled. 2. When transferring the cell mixture, the rectangular bag is prone to the situation where the liquid cannot be completely transferred to the cell culture bag, and too much liquid remains. Changing the rectangle into the convex-shaped bag in Example 2 can solve this problem well.

[0081] For the same fresh peripheral blood sample, the rectangular bag of Example 1 and the convex-shaped bag of Example 2 were used for culture under the same conditions for the same number of days. Comparison of various parameters showed that the convex-shaped bag was superior to the rectangular bag. See the comparison table below for details.

[0082] Further preferably, based on the same principle as above, in order to avoid wrinkles in the bag body and the formation of dead corners, which may lead to cell aggregation or residue, the corners of the primary cell storage bag 1 and the cell culture bag 4 are also designed to be rounded.

[0083] [Corrected 24.07.2024 according to Rule 91] In a further preferred embodiment, a quick-connector 8 is further added between pipes 505 and 507. As shown in Figures 7-13, the quick-connector 8 includes a male Luer connector 81 and a female Luer connector 82. Figure 7 is a schematic diagram of the structure of the male Luer connector 81, Figure 8 is a schematic diagram of the structure of the female Luer connector 82, and Figure 9 shows the male Luer connector 81 and the female Luer connector 82 being plugged together.

[0084] Specifically, referring to FIG7 , the male Luer connector 81 includes a first male mating portion 811, a first snap plate 812, a first gripping portion 813, and a first pipe connector 814. The first pipe connector 814 is used to connect to one of the pipelines 505 and 507. The first male mating portion 811 is inserted into the female mating portion of the female Luer connector 82 and the female Luer plug 84, described later, for mating connection. To facilitate communication with the connected pipelines 505 and 507, a first channel 815 is defined along the central axis of the first male mating portion 811, the first snap plate 812, the first gripping portion 813, and the first pipe connector 814.

[0085] The first clamping plate 812 is in the shape of a circular plate, and has at least two first notches 8121 disposed on its edge. Preferably, the two first notches 8121 are spaced apart, and more preferably, the two first notches 8121 are mirror-symmetrically disposed along the edge of the circular plate relative to its center. Furthermore, preferably, when there are three or more first notches 8121, they are uniformly spaced along the circumferential edge of the circular plate.

[0086] Preferably, the first pipe joint 814 is in a pagoda shape to facilitate sealing connection with the pipes 505 and 507.

[0087] 8 , the female Luer connector 82 includes a first female fitting portion 821, a first clamping portion 822, and a second pipe connector 823. The first female fitting portion 821 is provided with a first chamber 824 into which the first male fitting portion 811 is inserted. The second pipe connector 823 is provided with a second channel 825, which is in communication with the first chamber 824. The second pipe connector 823 is used to connect to another pipeline 505, 507.

[0088] Preferably, the first chamber 824 of the first female fitting portion 821 is cylindrical, one end of which is connected to the second pipe joint 823 and the other end is open.

[0089] The first clamping portion 822 of the female Luer connector 82 is provided on an end surface 8211 of the first female fitting portion 821 away from the second pipe connector 823 . The end surface 8211 is the end surface of the cavity wall at one end of the cylindrical first cavity 824 facing outward.

[0090] Furthermore, the first engaging portion 822 comprises at least two first engaging blocks 8221 having an L-shaped cross section, disposed on the end surface of the first female engaging portion 821. Preferably, the two first engaging blocks 8221 are spaced apart. Further preferably, the two first engaging blocks 8221 are disposed along the end surface 8211 of the first female engaging portion 821 in a circular mirror-symmetrical arrangement relative to the end surface. Further preferably, when there are three or more first engaging blocks 8221, the first engaging blocks 8221 are disposed on the end surface 8211 at uniform intervals.

[0091] Preferably, the second pipe joint 823 is in a pagoda shape to facilitate sealing connection with the pipes 505 and 507.

[0092] As shown in FIG9 , the first engaging plate 812 of the male Luer connector 81 and the first engaging portion 822 of the female Luer connector 82 are connected by engaging with each other through the first notch 8121 and the first engaging block 8221, thereby fixing the male Luer connector 81 and the female Luer connector 82 in a fixed connection and communicating the first channel 815 with the second channel 825.

[0093] Specifically, taking the two first notches 8121 and the two first latching blocks 8221 as an example, when connecting the male Luer connector 81 and the female Luer connector 82, the first male engaging portion 811 of the male Luer connector 81 is first inserted into the first female engaging portion 821 of the female Luer connector 82, and the first latching block 8221 of the female Luer connector 82 is aligned with the first notch 8121 of the male Luer connector 81. When the first latching block 8221 passes over the first notch 8121 along the connector axis, the male and / or female Luer connector is rotated through a certain angle, and the first latching block 8221 is engaged with the position of the first latching plate 812 where the first notch 8121 is not provided, thereby securing the male Luer connector 81 and the female Luer connector 82 together, as shown in FIG9. This connection structure allows for quick insertion, and does not require a large rotation angle to achieve a quick connection and fixation between the male and female Luer connectors 81 and 82.

[0094] Further preferably, at least one first annular groove 8111 is defined on the outer periphery of the first male mating portion 811. A first sealing ring (not shown) is disposed in the first annular groove 8111. Preferably, the first sealing ring protrudes from the outer surface of the first male mating portion 811 when disposed in the first annular groove 8111. The first sealing ring tightly engages with the inner sidewall of the first chamber 824 of the first female mating portion 821, thereby achieving a sealed connection between the male and female Luer connectors 81 and 82. Preferably, the first sealing ring is a silicone ring.

[0095] As an optional embodiment, a first sealing ring can be disposed on the end surface of the first male engaging portion 811 facing the female Luer connector 82. Thus, when the male Luer connector 81 and the female Luer connector 82 are mated, the first sealing ring is compressed between the bottom wall of the first chamber 824 of the female Luer connector 82 and the end surface of the first male engaging portion 811, thereby achieving a sealed connection between the male Luer connector 81 and the female Luer connector 82. Further preferably, the first sealing ring is disposed in an annular groove on the end surface of the first male engaging portion and protrudes from the surface of the end surface of the first male engaging portion.

[0096] Furthermore, as shown in FIG10 , the quick-plug connector 8 also includes a male Luer plug 83 that cooperates with the female Luer connector 82. The male Luer plug 83 includes a second male matching portion 831, a second clamping plate 832, and a second gripping portion 833. The second clamping plate 832 is provided with at least two second notches 8321 corresponding to the positions of the first clamping block 8221. The first clamping block 8221 can pass over the second notches 8321 and be clamped to the second clamping plate 832 after being rotated a certain angle to clamp the male Luer plug 83 and the female Luer connector 82 together, thereby achieving the goal of using the male Luer plug 83 to seal the female Luer connector 82 after the male Luer connector 81 and the female Luer connector 82 are disconnected, as shown in FIG11 .

[0097] Preferably, the male Luer plug 83 has no passage inside, or has a passage with one end closed inside.

[0098] Preferably, at least one second annular groove 8311 is provided on the outer periphery of the second male fitting part 831, and a second sealing ring (not shown in the figure) is provided in the second annular groove 8311. Preferably, when the second sealing ring is provided in the second annular groove 8311, it protrudes from the outer surface of the second male fitting part 831, and the second sealing ring is pressed into fit with the inner side wall of the first chamber 824 of the first female fitting part 821.

[0099] As an optional embodiment, a second sealing ring can be disposed on the end surface of the second male engaging portion 831 facing the female Luer connector 82. When the male Luer plug 83 is mated with the female Luer connector 82, the second sealing ring is compressed between the bottom wall of the first chamber 824 of the female Luer connector 82 and the end surface of the second male engaging portion 831, thereby achieving a sealed connection between the male Luer plug 83 and the female Luer connector 82 and sealing the female Luer connector 82. Further preferably, the second sealing ring is disposed in an annular groove on the end surface of the second male engaging portion and protrudes from the surface of the end surface of the second male engaging portion.

[0100] Furthermore, as shown in FIG12 , the quick-insert connector 8, 8' further includes a female Luer plug 84 that is mated with the male Luer connector 81. The female Luer plug 84 includes a second female mating portion 841 and a second clamping portion 842. The second clamping portion 842 is at least two second clamping blocks 8421 with an L-shaped cross-section provided on the end surface 8411 of the second female mating portion 841. The second clamping block 8421 can pass over the first notch 8121 and be clamped to the first clamping plate 812 after being rotated a certain angle, so as to clamp the female Luer plug 84 and the male Luer connector 81 together, and then the male Luer connector 81 is sealed with the female Luer plug 84, as shown in FIG13 .

[0101] Preferably, the second female fitting portion 841 is cylindrical and has a second chamber 843 inside. One end of the second chamber 843 is closed and the other end is open. The L-shaped second block 8421 is arranged on the end face 8411 of the open end of the second chamber 843 of the second female fitting portion 842.

[0102] Further preferably, the first and / or second snap-in plates 812 and / or 832 of the male Luer connector 81 have wedge-shaped or trapezoidal surfaces on the surfaces that engage the first and / or second L-shaped clamping blocks 8221 and / or 8421. Specifically, the thickness of the snap-in plates, facing the gripping portions 813 and 833, gradually increases from the first and second notches 8121 and 8321 toward the center between adjacent notches. Further preferably, the thickness remains constant between adjacent notches. This allows for a snap-in connection when the clamping blocks are moved across the notches and rotated a certain angle, allowing the snap-in connection and locking of the male and female Luer connectors 81 and 82 simultaneously due to the wedge-shaped or trapezoidal surfaces. Furthermore, when combined with embodiments in which the first and second sealing rings are positioned on the end surfaces of the male mating portion, the sealing rings can be compressed simultaneously with locking, further enhancing the sealing effect.

[0103] When the pipeline quick-plug connector 8 in the above embodiment connects the two sections of pipelines 505 and 505 and the pipelines 507 and 507, the first pipe connector of the male Luer connector 81 is connected to one end of the pipeline, and the female Luer connector 82 is connected to the other end of the pipeline. Then, the male Luer connector 81 and the female Luer connector 82 are quickly plugged and connected through the first clamping plate 812 and the first clamping portion 822 to achieve communication between the pipelines 505 and 507. The rotating clamping structure makes the connection fast and firm, improves the efficiency of liquid transfer, and can achieve a stable and tight connection between two different bag bodies through this quick plug-in clamping method. When there is no need to transfer liquid, the male Luer connector 81 and the female Luer connector 82 are disconnected, and then the male Luer plug 83 is quickly connected to the female Luer connector 82 to block the pipeline on the side of the female Luer connector 82 to prevent liquid leakage in the pipeline at this end of the female Luer connector 82. The female Luer plug 84 is quickly connected to the male Luer connector 81 to block the pipeline on the side of the male Luer connector 81 to prevent liquid leakage in the pipeline at this end of the male Luer connector 81.

[0104] Furthermore, another quick-plug connector 8' is provided at the end of the pipeline 506. The quick-plug connector 8' includes the above-mentioned female Luer connector 82 and the male Luer plug 83 connected thereto, see FIG11; or includes the above-mentioned male Luer connector 81 and the female Luer plug 84 connected thereto, see FIG13, to achieve rapid plugging of the pipeline 506.

[0105] During use, the cell culture bag set proposed in this patented invention requires only connecting the culture medium to the liquid inlet, injecting primary cells into the cell culture bag, and injecting the antibody coating solution into the initial cell activation bag to complete the preliminary preparations. The entire bag is placed in an automated culture workstation, and the tubing interconnects the bag to facilitate cell transfer and culture medium replenishment. This significantly reduces the number of manual steps and improves overall cell culture efficiency.

[0106] Obviously, those skilled in the art may make various changes and modifications to the embodiments of the present invention without departing from the spirit and scope of the present invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims and their equivalents, the present invention is intended to include such modifications and variations.

Claims

1. A fully automatic suspension cell culture bag set, characterized in that: The cell culture bag set includes: A primary cell storage bag, used to store primary cells, comprising a primary cell storage bag body, an injection port and a transfer port; An initial cell activation bag, used to activate initial suspended cells, comprising an initial cell activation bag body, a coating liquid injection port, a culture medium and cell liquid inlet port, and a liquid transfer port; A waste liquid bag, used for collecting waste liquid in the initial cell activation bag, the waste liquid bag comprising a waste liquid bag body and a liquid inlet; A cell culture bag, used for large-volume cell culture, comprising a cell culture bag body, a liquid inlet and a sampling port; Among them, the transfer outlet of the primary cell storage bag is connected to the culture medium and cell inlet of the initial cell activation bag through pipelines, and the liquid transfer outlet of the initial cell activation bag is connected to the waste liquid bag inlet and the cell culture bag inlet through pipelines.

2. The fully automatic suspension cell culture bag set according to claim 1, characterized in that: At least one of the injection port of the primary cell storage bag and the coating solution injection port of the initial cell activation bag is capped with sodium heparin.

3. The fully automatic suspension cell culture bag set according to claim 1, characterized in that: The cell culture bag also includes a cell density detection port, which is made of a light-transmitting material and can detect the cell density through the absorbance of the liquid in the culture bag.

4. The fully automatic suspension cell culture bag set according to claim 1, characterized in that: It also includes a four-way connector, three connectors of which are respectively connected to three culture medium inlet pipelines, and the fourth connector of the four-way connector is connected to the culture medium and cell inlet of the initial cell activation bag through pipelines.

5. The fully automatic suspension cell culture bag set according to claim 4, characterized in that: It also includes a three-way connector A, which is used to connect the pipeline connected to the transfer outlet of the primary cell storage bag, the pipeline connected to the fourth connector of the four-way connector, and the culture medium and cell inlet pipeline of the initial cell activation bag.

6. The fully automatic suspension cell culture bag set according to claim 5, characterized in that: It also includes a three-way connector B, which is used to connect the liquid transfer outlet pipeline of the initial cell activation bag, the liquid inlet pipeline of the waste liquid bag and the liquid inlet pipeline of the cell culture bag.

7. The fully automatic suspension cell culture bag set according to claim 6, characterized in that: The primary cell storage bag transfer outlet, the culture medium and cell inlet and liquid transfer outlet of the initial cell activation bag, and at least one of the liquid inlet of the cell culture bag are connected to the pipeline by a Luer connector.

8. The fully automatic suspension cell culture bag set according to any one of claims 1 to 7, characterized in that: The pipeline adopts a silicone tube.

9. The fully automatic suspension cell culture bag set according to any one of claims 1 to 7, characterized in that: At least one bag body among the primary cell storage bag, initial cell activation bag, waste liquid bag and cell culture bag is made of FEP waterproof and breathable film material.

10. The fully automatic suspension cell culture bag set according to any one of claims 1 to 7, characterized in that: The volume capacity of the cell culture bag is 2L or more.

11. The fully automatic suspension cell culture bag set according to claim 3, characterized in that: The cell density detection port is made of FEP material and comprises an upper light-transmitting hole and a lower light-transmitting hole which are opposed to each other, and the upper light-transmitting hole and the lower light-transmitting hole are blind holes.

12. The fully automatic suspension cell culture bag set according to any one of claims 1 to 7, characterized in that: Each corner of at least one of the primary cell storage bag, the initial cell activation bag and the cell culture bag is rounded.

13. The fully automatic suspension cell culture bag set according to claim 12, characterized in that: The initial cell activation bag is in the shape of a convex character with a narrow upper end and a wide lower end. The corners of the convex initial cell activation bag that bulge outward from the inner cavity are rounded. The coating liquid injection port, culture medium and cell liquid inlet and liquid transfer outlet are arranged at the upper end of the convex character.

14. The fully automatic suspension cell culture bag set according to claim 6, characterized in that: Also included is a quick-connect connector, including a male Luer connector and a female Luer connector that cooperates with the male Luer connector, wherein: The male Luer connector comprises a first male fitting portion, a first clamping plate, a first gripping portion, a first pipe connector, and a first passage penetrating the first male fitting portion, the first clamping plate, the first gripping portion, and the first pipe connector, wherein the first pipe connector is used to connect to a pipeline; The female Luer connector comprises a first female fitting portion, a first clamping portion, and a second pipe connector, wherein the first female fitting portion is provided with a first cavity into which the first male fitting portion is inserted, and the second pipe connector is provided with a second channel, the second channel being communicated with the first cavity, and the second pipe connector is used to be connected to a pipeline; The first clamping plate and the first clamping portion can be clamped and connected to fix the male Luer connector and the female Luer connector, and to connect the first channel with the second channel; The quick-insert connector is arranged on the pipeline between the four-way connector and the three-way connector A, and / or the pipeline between the three-way connector B and the liquid inlet of the cell culture bag, and / or the end of the medium liquid inlet pipeline.

15. The fully automatic suspension cell culture bag set according to claim 14, characterized in that: The first clamping portion is at least two first clamping blocks with L-shaped cross-sections arranged on the end surface of the first female matching portion. The first clamping plate is provided with at least two first notches corresponding to the positions of the first clamping blocks. The first clamping block can pass over the first notches and be clamped on the first clamping plate after being rotated a certain angle to clamp the male Luer connector and the female Luer connector together.

16. The fully automatic suspension cell culture bag set according to claim 14, characterized in that: At least one first annular groove is provided on the outer periphery of the first male matching part, a first sealing ring is provided in the first annular groove, and the first sealing ring is tightly fitted with the inner wall of the first cavity of the first female matching part; And / or, a first sealing ring is provided on the end surface of the first male fitting portion facing the female Luer connector, and the first sealing ring can be compressed between the bottom wall of the first chamber of the female Luer connector and the end surface of the first male fitting portion.

17. The fully automatic suspension cell culture bag set according to claim 15, characterized in that: It also includes a male Luer plug that is connected to the female Luer connector. The male Luer plug includes a second male matching portion, a second clamping plate and a second gripping portion. The second clamping plate is provided with at least two second notches corresponding to the position of the first clamping block. The first clamping block can pass over the second notches and be clamped on the second clamping plate after being rotated by a certain angle, so as to clamp the male Luer plug and the female Luer connector together.

18. The fully automatic suspension cell culture bag set according to claim 17, characterized in that: At least one second annular groove is provided on the outer periphery of the second male matching portion, a second sealing ring is provided in the second annular groove, and the second sealing ring is tightly fitted with the inner wall of the first cavity of the first female matching portion; And / or, a second sealing ring is provided on the end surface of the second male fitting portion facing the female Luer connector, and the second sealing ring can be compressed between the bottom wall of the first chamber of the female Luer connector and the end surface of the second male fitting portion.

19. The fully automatic suspension cell culture bag set according to claim 15, characterized in that: It also includes a female Luer plug that is connected to the male Luer connector. The female Luer plug includes a second female matching portion and a second clamping portion. The second clamping portion is at least two second clamping blocks with L-shaped cross-sections arranged on the end surface of the second female matching portion. The second clamping block can cross the first notch and be clamped on the first clamping plate after rotating a certain angle to clamp the female Luer plug and the male Luer connector together.

20. The fully automatic suspension cell culture bag set according to claim 15 or 19, characterized in that: A first clamping plate between adjacent first notches has a thickened wedge-shaped inclined surface or a trapezoidal inclined surface on one side facing the first gripping portion; And / or, a side of the second clamping plate between adjacent second notches facing the second gripping portion has a thickened wedge-shaped inclined surface or a trapezoidal inclined surface.

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