Culture bag device for small-scale culture of immune cells

Abstract

The utility model belongs to the technical field of biology, and particularly relates to a culture bag device for small-scale culture of immune cells. The utility model provides a culture bag device which comprises a bag body and a plurality of communicating pipelines, the bag body is hollow and internally provided with blocking faces, the blocking faces divide the bag body into a plurality of culture small chambers which are not communicated with one another, one end of each communicating pipeline is located in the corresponding culture small chamber, and the other end of each communicating pipeline is located in the corresponding culture small chamber. The other end of each communicating pipeline is located outside the bag body and provided with a connector assembly. The culture bag device disclosed by the utility model comprises six independent culture chambers, so that the requirement of developing a plurality of different process conditions on a small test scale of immune cells can be met, the potential risk of cell contamination caused by open type operation can be avoided, and good culture and amplification of the immune cells in a limited culture space can be realized.

Description

Technical Field

[0001] The utility model belongs to the field of biotechnology, and in particular relates to a culture bag device for small-scale culture of immune cells. Background Art

[0002] As the field of cell therapy matures, more and more companies are researching and developing various engineered cell products (such as CAR-T, TCR-T, CAR-NK, and TIL). Cell and Gene Therapy (CGT) products have become a frontier and hot topic in global pharmaceutical development.

[0003] The process development of cell therapy products includes multiple steps, such as donor peripheral blood / single blood collection, immune cell (T cell or NK cell, etc.) sorting, enrichment and activation, immune cell gene modification and in vitro expansion culture, and cell therapy product harvesting and freezing. The in vitro culture method of immune cells is the key to success. Currently, cell culture devices mainly include cell culture plates (such as 24-well plates and 6-well plates), cell culture flasks (such as T25 flasks and T75 flasks) and cell culture bags (such as 2L culture bags) for different scale cultures. Although cell culture plates and culture flasks can meet the needs of small-scale cell process development, they are open operations, which increases the risk of cell contamination. Therefore, the operator's awareness of aseptic operation is extremely high. Cell culture bags generally have a relatively large culture volume (such as 30ml-3L), and have the following advantages and characteristics: they can realize closed culture of immune cells, reducing the chance of cell contamination caused by open operations by operators; the material is transparent and flexible, which is convenient for direct observation of cell status under a microscope; the gas exchange area is large and the air permeability is good, which is conducive to large-scale culture in a limited culture space to obtain cell therapy products with the required dose for clinical transfusion; the bag material of the culture bag meets the pharmacopoeia resin pharmaceutical container standards and meets the clinical application requirements, etc. Therefore, cell culture bags are often used as the main expansion container for immune cell products. Based on the increasing demand for immune cell culture bags in the current cell therapy market and the scaledown linear amplification challenges faced by cell drug clinical applications from small-scale development of cell process to cell process amplification and locking, the development of a culture bag device that can be used for small-scale process development of immune cells is an urgent problem to be solved. The development of small-scale cell processes generally involves many experimental condition groups, and the lentiviral or retroviral transduction methods (static transduction or centrifugal transduction) used for different target gene modifications may be different. Although the existing 24-well or 6-well plates can achieve small-scale cell process development and meet the needs of differentiated transduction methods for different viruses, they cannot better linearly scale up the process development-related parameters to the cell culture bag, and are manual open operations, with the potential risk of cell contamination due to open operations. In order to simultaneously meet the needs of small-scale cell amplification and differentiated transduction methods for different viruses, and at the same time match the adapter requirements of high-end centrifuges of the same brand in the cell therapy market during cell centrifugal transduction, and better reflect the scaledown linear amplification model of cell drugs from small-scale process development to process amplification locking, while avoiding the potential risk of cell contamination due to open operations, it is imperative to develop a new culture bag device that can be used for small-scale culture of immune cells. Utility Model Content

[0004] In order to solve the above-mentioned defects of the prior art, the utility model provides a culture bag device for small-scale culture of immune cells.

[0005] In a specific embodiment of the present invention, the culture bag device includes a bag body and a plurality of connecting pipes. The bag body is hollow and has a barrier surface disposed therein. The barrier surface divides the bag body into a plurality of mutually disconnected culture chambers. One end of each connecting pipe is located within and communicates with a corresponding culture chamber, and the other end of each connecting pipe is located outside the bag body and is provided with an interface assembly. The bag body is used to culture immune cells (such as T cells and NK cells), the culture chambers are used to culture a specific type of immune cell under different culture conditions, and the interface assembly is used to seal the culture chambers and connect them to a syringe.

[0006] In a specific embodiment of the present invention, the barrier surface includes a first barrier surface and a plurality of second barrier surfaces, and the bag body, the first barrier surface and the plurality of second barrier surfaces are integrally formed.

[0007] In a specific embodiment of the present invention, the outer peripheral surfaces of the first barrier surfaces are all connected to the inner wall of the bag body, the outer peripheral surfaces of several second barrier surfaces are all connected to the inner wall of the bag body, several second barrier surfaces are cross-connected to the first barrier surface, and the spacing distances between adjacent second barrier surfaces on the first barrier surface are equal, that is, the distance between the second barrier surface and the bag body and the distance between any two barrier surfaces are equal, so that the first barrier surface and the second barrier surface can divide the bag body into completely independent and non-interfering culture chambers.

[0008] In a specific embodiment of the present invention, a plurality of the second blocking surfaces are perpendicular to the first blocking surface.

[0009] In a specific embodiment of the present invention, the number of the second barrier surfaces is at least two, for example, three, four, five, six or more.

[0010] In a preferred embodiment of the present invention, the number of the second barrier surfaces is two.

[0011] In a specific embodiment of the present invention, each of the culture chambers does not interfere with each other.

[0012] In a specific embodiment of the present invention, the number of the culture chambers is at least six; for example, it can be six, eight, or ten or more.

[0013] In a preferred embodiment of the present invention, the number of the culture chambers is six.

[0014] In a specific embodiment of the present invention, the number of the connecting pipes is equal to the number of the culture chambers.

[0015] In a specific embodiment of the present invention, the interface assembly includes a female Luer interface and a male Luer cap. The female Luer interface is clamped on the end of the connecting pipeline away from the bag body, and the male Luer cap is detachably connected to the female Luer interface.

[0016] In a preferred embodiment of the present invention, the male Luer cap is threadedly connected to the female Luer interface.

[0017] In a specific embodiment of the present invention, a clamp is further provided on the connecting pipeline between the female Luer interface and the bag body for opening or closing the connecting pipeline.

[0018] In a specific embodiment of the present invention, the bag body is made of a transparent, breathable material; the barrier surface is made of an airtight material.

[0019] The beneficial effects of the present invention are:

[0020] Compared to the commonly used ordinary six-well plates, the six-row culture bag for small-scale culture of immune cells of the present invention completes the entry or removal of raw materials for each process in the small-scale process development of immune cells through a closed connection with a screw-mouth syringe, which can avoid the potential risk of cell contamination caused by open operation. The good air permeability of the surface of the six-row culture bag can better achieve good gas exchange between immune cells and the outside world, and achieve good culture and expansion of immune cells in a limited culture space. Currently, there are no multi-chamber culture bags for small-scale process development of immune cells on the market. The six-row culture bag of the present invention contains six separate culture chambers that can meet the development needs of multiple different process conditions for small-scale immune cells, and the developed process parameters can achieve scaledown linear amplification to large-volume cell culture bags for scaled-up production. In addition, the six-row culture bag can match the adapter requirements of the centrifuge during centrifugal transduction of immune cells, and realize cell centrifugal transduction operations through the culture bag. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a structural diagram of the culture bag device of the present utility model.

[0022] Figure 2 It is a side view of the culture bag device of the present invention.

[0023] Reference numerals:

[0024] 1-connecting pipeline; 2-interface assembly; 21-female Luer interface; 22-male Luer cap; 3-bag body; 4-clamp; 5-culture chamber; 6-barrier surface; 61-first barrier surface; 62-second barrier surface. DETAILED DESCRIPTION

[0025] In the description of the present invention, it should be noted that the structures, proportions, sizes, etc. illustrated in the drawings of this specification are only used to match the contents disclosed in the specification for understanding and reading by those familiar with the art, and are not intended to limit the conditions for the implementation of the present invention. Therefore, they have no technical significance. Any structural modifications, changes in proportional relationships, or adjustments in size, without affecting the efficacy and objectives of the present invention, should still fall within the scope of the technical content disclosed in the present invention. At the same time, the terms "center", "longitudinal", "transverse", "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc., indicating directions or positional relationships, are based on the directions or positional relationships shown in the drawings and are only for the convenience of describing the present invention and simplifying the description. They do not indicate or imply that the devices or components referred to must have a specific orientation, be constructed and operate in a specific orientation, and therefore should not be understood as limiting the present invention. In addition, the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance.

[0026] In the description of this utility model, it should be noted that, unless otherwise expressly specified or limited, the terms "installed," "connected," "connected," and "connected" should be understood in a broad sense. For example, they may refer to fixed connections, detachable connections, or integral connections; mechanical connections or electrical connections; direct connections or indirect connections through an intermediate medium; or internal communication between two components. Those skilled in the art will be able to understand the specific meanings of the above terms in this utility model based on specific circumstances.

[0027] The utility model provides a culture bag device for small-scale culture of immune cells, as shown in the attached Figure 1 shown.

[0028] The culture bag device includes a bag body 3, a bag-shaped structure. Made of materials that meet pharmacopoeial standards for resin pharmaceutical containers, the bag body 3 is transparent and flexible, facilitating direct microscopic observation of cell status. Furthermore, the bag body 3 exhibits excellent air permeability, enabling efficient gas exchange between immune cells and the environment. The bag body 3 is constructed from EVA (ethylene-vinyl acetate copolymer), COC / LLDPE (cyclic olefin copolymer / low-density polyethylene), EVA / LDPE (ethylene-vinyl acetate copolymer / low-density polyethylene), or a medical-grade thermoplastic elastomer.

[0029] The bag body 3 is hollow and contains an airtight barrier surface 6. The barrier surface 6 is made of polyvinyl chloride, polyethylene, polypropylene, polyvinylidene fluoride, rubber, or an elastomeric material. The barrier surface 6 divides the bag body 3 into six culture chambers 5, each of equal volume. The barrier surface 6 includes a first barrier surface 61 and two second barrier surfaces 62, which are integrally formed. The first barrier surface 61 is arranged horizontally, while the two second barrier surfaces 62 are arranged longitudinally. The outer periphery of the first barrier surface 61 is connected to the inner wall of the bag body 3, while the outer periphery of the two second barrier surfaces 62 is connected to the inner wall of the bag body 3. The two second barrier surfaces 62 are cross-connected to the first barrier surface 61, and the second barrier surfaces 62 are spaced equidistantly on the first barrier surface 61. That is, the distance between the second barrier surface 62 and the bag body 3 and the distance between each second barrier surface 62 are equal. The first blocking surface 61 is vertically disposed in the middle of the second blocking surface 62 .

[0030] The bag 3 is divided into mutually exclusive culture chambers 5 by a barrier surface 6. Each culture chamber 5 can have equal or unequal volumes. Within each culture chamber 5, immune cells of a specific type can be cultured separately under different culture conditions, without interfering with each other's culture. Each culture chamber 5 is an independent, enclosed space, with the exception of the connecting pipe 1, which serves as a single inlet and outlet.

[0031] The culture bag device also includes six connecting pipes 1, which correspond one to one with the six culture chambers 5. One end of each connecting pipe 1 is located in the corresponding culture chamber 5 and is connected to the corresponding culture chamber. The other end of each connecting pipe is located outside the bag body and is provided with an interface component, thereby realizing the connection between the culture chamber 5 and the syringe. Raw materials such as cell fluid, culture medium and virus enter the culture chamber 5 and are taken out of the culture chamber 5 through the connecting pipe 1 during the cell culture process.

[0032] Each connecting line 1 is provided with an interface assembly 2 at one end away from the bag body 3. The interface assembly 2 can be a conventional interface assembly used in cell culture, such as a Luer interface, a bayonet interface, a sterile interface, a screw interface, a crimp interface, a quick-connect interface, a quick-plug design, a needle interface, an O-ring interface, a septum interface, or a magnetic interface. The interface assembly 2 ensures that the culture chamber 5 is sealed during cell culture and can be connected to a sterile screw-thread syringe or other device in a closed manner. During the cell culture process, the sterile screw-thread syringe connected to the interface assembly 2 is used to inject raw materials required for cell culture into the culture chamber 5, and to remove waste liquid generated by cell culture from the culture chamber 5.

[0033] Specifically, the interface assembly 2 includes a female Luer interface 21 and a male Luer cap 22. The female Luer interface 21 is snap-fitted to the end of the connecting pipe 1 away from the bag body 3, and the male Luer cap 22 is threadedly connected to the female Luer interface 21. The female Luer interface 21 is used to connect to a sterile screw-cap syringe with a male Luer interface for injecting and removing raw materials such as cell fluid, culture medium, and viruses during cell culture. The male Luer cap 22 is used to seal the female Luer interface 21 during cell culture. When the male Luer cap 22 is connected to the female Luer interface 21 and tightened and locked, the interface assembly 2 forms a closed structure to prevent external contamination from entering the culture chamber 5.

[0034] A clamp 4 is also provided on the connecting pipe 1 between the female Luer connector 21 and the bag body 3. The clamp 4 can be switched between an open and a closed state, allowing the entire connecting pipe 1 to be opened or closed. When raw materials need to pass through the connecting pipe 1, the clamp 4 is opened. After the raw materials are injected or removed, the clamp 4 is closed to seal the culture bag assembly.

[0035] To use the culture bag device of the present invention, the following steps are performed: The culture bag device is placed in a biosafety cabinet for aseptic operation. The male Luer cap 22 is removed from the female Luer connector 21 by rotating it. A sterile screw-capped syringe containing cell fluid, culture medium, or other raw materials and having a male Luer connector is connected to the female Luer connector 21. The clamp 4 is opened, and the cell fluid, culture medium, or other raw materials in the sterile screw-capped syringe are injected into the culture chamber 5. After the injection is completed, the sterile screw-capped syringe with the male Luer connector is removed from the female Luer connector 21, the male Luer cap 22 is replaced and tightened, and the clamp 4 is closed. The culture bag device is then laid flat in a cell culture incubator under suitable cell growth conditions for incubation.

[0036] When fluid replacement or sampling is required, the male Luer cap 22 is removed from the female Luer port 21 by rotating it. A sterile screw-capped syringe with a male Luer port is connected to the female Luer port 21. The clamp 4 is opened and the old culture medium is slowly aspirated with the sterile screw-capped syringe. After aspiration is complete, the sterile screw-capped syringe with a male Luer port containing the old culture medium is removed from the female Luer port 21. A sterile screw-capped syringe with a male Luer port containing fresh culture medium is connected to the female Luer port 21. The plunger of the sterile screw-capped syringe is pushed to inject the fresh culture medium into the culture chamber 5. After injection is complete, the sterile screw-capped syringe with a male Luer port is removed from the female Luer port 21, the male Luer cap 22 is replaced and tightened, and the clamp 4 is closed. The culture bag assembly is laid flat in a cell culture incubator under conditions suitable for cell growth.

[0037] The above embodiments are merely illustrative of the principles and effects of the present invention and are not intended to limit the present invention. Anyone skilled in the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by one of ordinary skill in the art without departing from the spirit and technical principles disclosed in the present invention are intended to be covered by the claims of the present invention.

Claims

1. A culture bag device for small-scale culture of immune cells, characterized in that: The culture bag device comprises a bag body (3) and a plurality of connecting pipes (1); the bag body (3) is hollow and has a barrier surface (6) arranged therein; the barrier surface (6) divides the bag body (3) into a plurality of mutually unconnected culture chambers (5); one end of each connecting pipe (1) is located in the corresponding culture chamber (5) and is connected to the corresponding culture chamber (5); the other end of each connecting pipe (1) is located outside the bag body (3) and is provided with an interface assembly (2).

2. The culture bag device according to claim 1, characterized in that: The barrier surface (6) comprises a first barrier surface (61) and a plurality of second barrier surfaces (62); the bag body (3), the first barrier surface (61) and the plurality of second barrier surfaces (62) are integrally formed.

3. The culture bag device according to claim 2, characterized in that: The outer peripheral surfaces of the first barrier surface (61) are all connected to the inner wall of the bag body (3), the outer peripheral surfaces of a plurality of the second barrier surfaces (62) are all connected to the inner wall of the bag body (3), a plurality of the second barrier surfaces (62) are cross-connected to the first barrier surface (61), and the spacing distances between adjacent second barrier surfaces (62) on the first barrier surface (61) are equal.

4. The culture bag device according to claim 3, characterized in that: A plurality of the second blocking surfaces (62) are perpendicular to the first blocking surface (61).

5. The culture bag device according to any one of claims 3 or 4, characterized in that: The number of the second blocking surfaces (62) is at least two.

6. The culture bag device according to claim 5, characterized in that: The number of the second blocking surfaces (62) is two.

7. The culture bag device according to claim 1, characterized in that: Each of the culture chambers (5) does not interfere with each other.

8. The culture bag device according to claim 1, characterized in that: The number of the culture chambers (5) is at least six; And / or, the number of the connecting pipes (1) is equal to the number of the culture chambers (5).

9. The culture bag device according to claim 8, characterized in that: The number of the culture chambers (5) is six.

10. The culture bag device according to claim 1, characterized in that: The interface assembly (2) comprises a female Luer interface (21) and a male Luer cap (22); the female Luer interface (21) is snap-connected to an end of the connecting pipe (1) away from the bag body (3); and the male Luer cap (22) is detachably connected to the female Luer interface (21).

11. The culture bag device according to claim 10, characterized in that: The male Luer cap (22) is threadedly connected to the female Luer interface (21).

12. The culture bag device according to claim 11, characterized in that: A clamp (4) is also provided on the connecting pipe (1) between the female Luer interface (21) and the bag body (3).

13. The culture bag device according to claim 1, wherein: The bag body (3) is made of a transparent, breathable material; and / or the barrier surface (6) is made of an airtight material.

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