Cell culture bag

Abstract

The utility model discloses a cell culture bag which structurally comprises a body, at least two cavities used for containing cell culture fluid are arranged on the body, a partition strip capable of being broken through by extruding the cavities is arranged between every two adjacent cavities, and the cavity located at the head end is provided with an input port used for injecting original cells. And the cavity at the tail end is provided with an output port for outputting cell culture fluid after culture and amplification. According to the CAR-T cell culture bag, culture media are preloaded in advance through the independent cavities, when the CAR-T cell culture bag is used, only the separation belts between the cavities need to be extruded, the next cavity preloaded with the culture media is communicated with the previous cavity, and replacement of the culture bag and replacement of liquid in the CAR-T cell whole culture period can be avoided; therefore, pollution caused by frequent replacement of the culture bag and in the liquid supplementing process is avoided.

Description

Technical Field

[0001] The utility model relates to a bag-type structure, in particular to a cell culture bag. Background Art

[0002] CAR-T therapy is chimeric antigen receptor T cell immunotherapy, a new type of precision targeted therapy for treating tumors. It uses genetic engineering technology to transform and culture immune cells collected from the patient in vitro and then return them to the patient to stimulate and enhance the body's own immune function, thereby achieving the purpose of treating tumors.

[0003] At present, CAR-T cells are generally expanded at high density through cell culture bags. The gas exchange area of ​​the cell culture bag is large, which is conducive to the culture of cells, and the cell culture bag is convenient for microscopic observation and monitoring of the cell culture process. However, during the overall culture and expansion process, multiple rehydration is required, which is cumbersome to operate, which greatly increases the chance of cell contamination; and when the number of cells reaches a certain level, the cells need to be transferred to a larger culture bag to expand the volume of culture.

[0004] Existing cell culture bags are not convenient for cell transfer, the cell transfer operation is complicated, and liquid residue is prone to occur. Patent 220132239U discloses a convenient and safe immune cell culture bag. After the cell culture begins, this patent will still be connected to the outside world through various pipes, and cannot guarantee the sterility and airtightness during the entire culture cycle. There is still a great risk of bacterial contamination. Therefore, existing cell culture bags have problems such as complex rehydration operation, numerous devices, inability to continuously guarantee the sterility and airtightness of the cell culture system, and easy cell contamination. Utility Model Content

[0005] Purpose of the utility model: The purpose of the utility model is to provide a cell culture bag that does not require replacement of culture bags and liquids and reduces cell contamination.

[0006] Technical solution: The cell culture bag described in the utility model includes a main body, which is provided with at least two cavities for containing cell culture fluid, and a dividing zone that can be broken by squeezing the cavity is provided between adjacent cavities. The cavity located at the head end is provided with an input port for injecting original cells, and the cavity located at the tail end is provided with an output port for outputting the culture fluid of the cells after culture and expansion.

[0007] Among them, the cell culture bag described in the utility model is a CAR-T cell full-cycle culture bag; the volume of the cavity close to the output port is greater than or equal to the volume of the cavity close to the input port; more preferably, the volume of each cavity increases from the head end to the tail end; or, when the number of cavities is greater than or equal to 3, the volume of the first cavity is the same as that of the second cavity, and from the third cavity onwards, the volume of each cavity is larger than that of the previous cavity.

[0008] The sealing property of the left side of the separation belt is greater than that of the right side of the separation belt, so that the separation belt can be broken only when the cavity near the output port is squeezed toward the cavity near the input port; the separation belt is preferably a hot pressing belt.

[0009] Wherein, the input port and the output port are both provided with a detachable sealing structure; the sealing structure includes a neck connected to the head end cavity or the tail end cavity and a cover body for sealing the neck; the neck is inclined at an angle to the cavity; the cover body includes a top cover and a bottom cover; an elastomer is provided between the top cover and the bottom cover to facilitate the syringe to penetrate the bottom cover and thus connect with the head end cavity or the tail end cavity; the elastomer is an injection point provided on the surface of the bottom cover for easy acupuncture; the injection point is circular in shape, the number of circular injection points is three, one of the three circular injection points is a larger circle, and the other two are smaller circles; the elastomer is rubber.

[0010] Among them, the top cover and the bottom cover, and the bottom cover and the neck are respectively threadedly connected; the bottom cover is provided with a bottom cover threaded portion protruding upward and a bottom cover lower edge extending downward along the bottom cover for threaded connection with the neck, and the bottom cover threaded portion has both external threads and internal threads, the external threads are used for threaded connection with the top cover 321, and the internal threads are used for threaded connection with the neck; the top cover is provided with a top cover lower edge extending downward along the top cover connected to the bottom cover threaded portion; the top cover is provided with a protruding portion.

[0011] Beneficial effects: Compared with the prior art, the utility model has achieved the following significant effects: (1) The bag body is divided into multiple independent cavities by a separation belt that can be broken through by squeezing, and a culture medium delivery port is provided at both ends. By pre-dividing the culture medium in advance, the culture bag and the liquid do not need to be replaced during the entire culture cycle of CAR-T cells, thereby reducing the pollution caused by replacing the culture bag and replenishing the liquid; (2) The volume of each cavity is cleverly set to increase in sequence from the input port to the output port, meeting the requirement that the culture medium required in the late stage of the cell culture cycle is more than the culture medium required in the early stage; (3) The sealing of the left side of the separation belt is greater than the sealing of the right side of the separation belt, so that the separation belt can be broken only when the cavity near the output port is squeezed toward the cavity near the input port, thereby avoiding squeezing of cells. The cell side will cause damage to the cells, and it can also prevent premature culture medium expansion caused by mispressure during the initial cell infusion. Therefore, it not only ensures that the cells are not damaged but also prevents the risk of operational errors; (4) The double-layer cover structure can take into account a variety of ways to add initial cells. When it is necessary to use a syringe to inject the original cells, you only need to unscrew the first layer of the top cover, insert the syringe into the rubber port and inject the cells. There is no need to open the second layer of the bottle cap. If other methods are used to add original CAR-T cells, unscrew the second layer of the bottom cover and add them from the opening of the neck. This can comprehensively reduce the risk of cell contamination during the process; (5) The utility model is an integrated product with a simple structure and high practicality. It can comprehensively solve the problems of complex operation and numerous devices in the process of CAR-T cell culture. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 It is a schematic diagram of the top view structure of the utility model;

[0013] Figure 2 This is a schematic diagram of the side view structure of the utility model after being filled with cell culture fluid;

[0014] Figure 3 This is a schematic diagram of the top cover structure of the utility model;

[0015] Figure 4 It is a schematic diagram of the bottom cover structure of the utility model. DETAILED DESCRIPTION

[0016] The technical solution of the utility model is further described below in conjunction with the accompanying drawings of the specification.

[0017] like Figure 1-2As shown, the utility model provides a cell culture bag, including a body 2, on which at least two cavities 21 for containing cell culture fluid are provided, and a separation belt 22 that can be broken by squeezing the cavity is provided between adjacent cavities, and the separation belt 22 is a hot pressing belt; the cavity at the head end is provided with an input port 1 for injecting original cells, and the cavity at the tail end is provided with an output port 3 for outputting the culture fluid of the cultured and amplified cells. The volume of the cavity 21 close to the output port is greater than or equal to the volume of the cavity close to the input port 1; more preferably, the volume of each cavity increases in sequence from the head end to the tail end; or, when the number of cavities is greater than or equal to 3, the volume of the first grid cavity is the same as that of the second grid cavity, and from the third grid onwards, the volume of each grid cavity is larger than that of the previous grid cavity. When specifically applied to CAR-T cell culture, since the culture and expansion cycle is generally 8-10 days, during which 4-6 rehydration is required, the utility model divides the culture bag into 6 compartments, and the volume ratio of the six compartment cavities 21 is 1:1:2:4:8:16, and other ratios can also be set as needed; Figure 1 The cavity in the figure only represents the cavity structure, and does not represent the actual volume size and relative volume size of each cavity. The sealing performance of the left side of the separation zone 22 is greater than the sealing performance of the right side of the separation zone 22, and the separation zone 22 can be broken when the cavity near the output port 3 is squeezed toward the cavity near the input port 1.

[0018] Both the input port 1 and the output port 3 are provided with a detachable sealing structure. The sealing structures of the input port 1 and the output port 3 of this embodiment are the same, so only the sealing structure of the output port 3 is taken as an example for detailed description. The sealing structure of the output port 3 includes a neck 31 connected to the tail end cavity and a cover 32 for sealing the neck 31. Similarly, the sealing structure of the input port 1 includes a neck connected to the head end cavity and a cover for sealing the neck. Figure 2 As shown, in the longitudinal section, the two necks are inclined with the cavity respectively. Therefore, the input port 1 and the output port 3 of the present application are both provided with inclined neck structures, which can effectively prevent the culture solution in the cavity 21 from overflowing.

[0019] like Figure 3 , 4As shown, the cover body 32 includes a top cover 321 and a bottom cover 322; the top cover 321 and the bottom cover 322, and the bottom cover 322 and the neck 31 are respectively threadedly connected; the bottom cover 322 is provided with a bottom cover threaded portion 3221 protruding upward and a bottom cover lower edge extending downward along the bottom cover 322 for threaded connection with the neck 31, and the bottom cover threaded portion has both external threads and internal threads, the external threads are used for threaded connection with the top cover 321, and the internal threads are used for threaded connection with the neck; the top cover 321 is provided with a lower edge of the top cover 321 extending downward along the top cover 321 for threaded connection with the bottom cover threaded portion 3221, and the top cover is provided with an upward protrusion. An elastic body 3222 is provided between the top cover 321 and the bottom cover 322 to facilitate acupuncture injection; the elastic body 2222 is an injection point provided on the surface of the bottom cover 322 to facilitate acupuncture. The injection points of this embodiment are circular, and there are three of them. One of the three circular injection points is a larger circle, and the other two are smaller circles. The material of these three injection points is rubber.

[0020] Since the culture and expansion cycle of CAR-T cells is generally 8-10 days, 4-6 times of rehydration are required during this period. The utility model divides the culture bag into 6 grids. When in use, an appropriate amount of culture medium is pre-installed in each grid in advance. Since the total amount of cells is more than the previous time each time the rehydration is performed, the amount of each rehydration will also increase compared to the previous time. Therefore, the volume of the 6 grids of the culture bag is the same as that of the first and second grids, and the volume increases step by step from the third grid to ensure that sufficient culture fluid can be replenished each time. The dividing strip 22 between adjacent cavities 21 is unidirectionally extruded and can pass through, that is, it can only be squeezed from the cavity near the output port 3 to the cavity near the input port 1, that is, when the culture medium needs to be supplemented, the culture medium can only be squeezed to break through the partition by squeezing the side of the culture medium and merged with the cell fluid already in culture, so that not only can the damage to the cells caused by squeezing the cell side be avoided, but also the early expansion of the culture medium caused by mispressure during the initial cell infusion can be prevented, thereby ensuring that the cells are not damaged and preventing the risk of operational errors.

[0021] The cover 32 of the utility model adopts a double-layer structure. After the top cover 321 on the first layer is unscrewed, a rubber sealing port is provided on the surface of the bottom cover 322. When the experimenter wants to inject the original cells by needle, he only needs to unscrew the first top cover 321, insert the needle into the rubber port and inject the cells. There is no need to open the second bottle cap, which greatly reduces the contact with the outside world and reduces the possibility of contamination. If other methods are used to add original CAR-T cells, unscrew the bottom cover 322 of the second layer and add them from the opening of the neck 31.

Claims

1. A cell culture bag, characterized in that: The invention comprises a main body (2), wherein the main body (2) is provided with at least two cavities (21) for containing cell culture fluid, a separation zone (22) which can be broken through by squeezing the cavities (21) is provided between adjacent cavities (21), the cavity at the head end is provided with an input port (1) for injecting original cells, and the cavity at the tail end is provided with an output port (3) for outputting culture fluid of cells after culture and expansion.

2. The cell culture bag according to claim 1, characterized in that: The volume of the cavity (21) on the side close to the output port is greater than or equal to the volume of the cavity on the side close to the input port (1).

3. The cell culture bag according to claim 1, characterized in that: The sealing performance of the left side of the separation belt (22) is greater than the sealing performance of the right side of the separation belt (22), so that the separation belt (22) can be broken when the cavity near the output port (3) is squeezed toward the cavity near the input port (1).

4. The cell culture bag according to claim 1, characterized in that: The input port (1) and the output port (3) are both provided with a detachable sealing structure.

5. The cell culture bag according to claim 4, characterized in that: The sealing structure comprises a neck connected with the head end cavity or the tail end cavity and a cover body used for sealing the neck.

6. The cell culture bag according to claim 5, characterized in that: The cover body comprises a top cover and a bottom cover; an elastic body is arranged between the top cover and the bottom cover to facilitate the syringe to penetrate into the bottom cover so as to communicate with the head end cavity or the tail end cavity.

7. The cell culture bag according to claim 6, characterized in that: The top cover and the bottom cover, and the bottom cover and the neck are respectively threadedly connected; the bottom cover is provided with a bottom cover threaded portion protruding upward and a bottom cover lower edge extending downward along the bottom cover for being threadedly connected to the neck; the top cover is provided with a top cover lower edge connected to the bottom cover threaded portion and extending downward along the top cover.

8. The cell culture bag according to claim 6, characterized in that: The elastic body is an injection point arranged on the surface of the bottom cover for easy needle puncture.

9. The cell culture bag according to claim 5, characterized in that: The neck is inclined with respect to the cavity.

10. The cell culture bag according to claim 1, characterized in that: The separation belt (22) is a hot pressing belt.

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