CN204417517U - A kind of mass cell cultivates multiple-bag - Google Patents

Abstract

The utility model relates to the technical fields of biomedical engineering and medical inspection, in particular to a large-scale cell culture multi-pack, comprising a culture bag body, a sample bag system and a collection bag system, one side of the culture bag body is connected to the sample bag in sequence Connection port, collection bag connection port, raw material sampling port, supplementary sampling port and liquid replacement filter port, the sample bag system includes a sample bag connecting tube and a sample bag connected to the sample bag connecting tube, the sample bag connecting port connected to the sample bag connecting pipe; the collecting bag system includes a collecting bag connecting pipe and a collecting bag connected to the collecting bag connecting pipe. The beneficial effects of the utility model are that the structure is simple, the operation is convenient, good stability, high reliability, and convenient operation, which is beneficial to the cultivation of cells and obtaining more high-quality cells, and can directly perform various operations such as microscopic observation. , which can realize cell expansion, sampling and collection in a closed system, improve cell culture efficiency and reduce contamination rate.

Description

A large-scale cell culture multi-pack

technical field

The utility model relates to the technical fields of biomedical engineering and medical inspection, in particular to a large-scale cell culture multiple bag.

Background technique

Cell culture generally refers to the cultivation of tissues or cells taken from living animals in vitro and simulates the physiological conditions in vivo to achieve continuous growth and expansion of cells. Cell culture is the basic condition for cell growth and is widely used in the fields of biology, medicine and pharmacy. When cell culture is carried out in the current scientific research field, PET or PET6 material culture bottles are mostly used, which are used after being sterilized by Gamma rays, which are disposable consumables. However, since the culture bottle belongs to an open system, it cannot realize a fully closed state, fails to reach the GMP standard, and does not meet the clinical treatment norms. The application of cell culture bags can realize a closed cell culture system, which can provide a reliable source of cells for clinical use in GMP laboratories.

The Chinese utility model patent with the application number "201320010782.8", the authorized announcement number "CN 203112848 U", and the title "a polygonal plastic culture bag for cell culture" discloses a polygonal cell culture bag, which consists of: The bottom sheet of the plastic bag, the bottom sheet of the polygonal plastic bag is welded with the top sheet of the plastic bag, the top sheet of the plastic bag has an air inlet and a feeding port, and the bottom sheet of the polygonal plastic bag is a regular hexagon, a regular octagon or a regular decagon quadrilateral. The defect of this utility model patent is: there is no sample retention bag, and the quantity and quality of the cells in the culture bag cannot be monitored; there is no cell collection bag, and the cultured cells can only be collected at one time; there is no liquid replacement system, and the cultured cells cannot be changed and Supplement with new medium.

The Chinese invention patent with the application number "200980110585.X", the authorized announcement number "CN 101978041 B", and the title "Multilayer Cell Culture System with Restricted Passage" discloses a multilayer cell culture device, which includes: a . at least three cell culture chambers, each cell culture chamber having at least one port; b. at least two integral catheter chambers; and c. a sliding port cover constructed and arranged to interface with the multilayer cell culture chamber; At least one port of at least three cell culture chambers of the device is engaged and an open or closed port is provided, wherein the open or closed channel is determined by placing the port cover in an open position relative to the at least one port Or closed position sliding engagement, wherein the open position port cover includes a filter and the closed position port cover includes a septum; the sliding port cover is connected to the multilayer cell culture device by a hinged connector. The disadvantages of this invention patent are: complex structure, high product cost, non-disposable consumables, suitable for the culture of adherent cells, and not suitable for the needs of clinical cell therapy.

Utility model content

The purpose of the utility model is to overcome the deficiencies of the prior art, and provide a fully enclosed multi-pack for large-scale cell culture and collection, which is scientific and reasonable, convenient to use, and good in cultivating effect.

In order to achieve the above purpose, the utility model provides a large-scale cell culture multi-package bag, including a culture bag body, a sample bag system and a collection bag system, one side of the culture bag body is connected to the connection port of the sample bag, the collection bag Connecting port, raw material sampling port, supplementary sampling port and liquid replacement filter port, of which:

The sample bag system includes a sample bag connecting pipe and a sample bag connected to the sample bag connecting pipe, and the sample bag connecting port communicates with the sample bag connecting pipe;

The collection bag system includes a collection bag connection pipe and a collection bag connected to the collection bag connection pipe, and the collection bag connection port communicates with the collection bag connection pipe.

Preferably, the raw material sampling port connects the raw material cell bag and the culture bag through a sterile adapter, and transfers the cells to be cultured into the cell culture bag body.

Preferably, a first dropping funnel protrudes from the feeding port, and a first dropping funnel plug is sleeved on the first dropping funnel;

The first dropping funnel and the first dropping funnel plug are detachable structures.

Preferably, the first dropping funnel on the feeding port is connected to a syringe to add nutritional factors or cytokines to the cell culture bag body.

Preferably, a second dropping funnel protrudes from the filter port for changing liquid, and a second dropping funnel plug is sleeved on the second dropping funnel;

Two layers of filter membranes are sequentially arranged in the second dropping funnel along its axial direction;

The second dropping funnel and the second dropping funnel plug are detachable structures.

Preferably, the filter membrane in the second dropping funnel close to the second dropping funnel plug is a 70um cell filtration membrane; the filter membrane in the second dropping funnel away from the second dropping funnel plug is a 3um cell filtration membrane.

Preferably, the second dropping funnel on the filter port for changing the liquid is connected to a negative pressure suction device to realize long-term culture and liquid replacement in the cell culture bag body.

Preferably, the 70um cell filtration membrane is a nylon membrane to prevent large cell aggregates from clogging the filtration system.

Preferably, the 3um cell filter membrane is a transwell membrane, which is used to prevent the cultured cells from being sucked away during negative pressure suction.

Preferably, the collection bag is provided with a collection bag test sample bag;

The collection bag and the collection bag test sample bag are connected through the test sample bag connecting pipe.

Preferably, the culture bag body is a one-piece structure.

Preferably, the culture bag body adopts a heat sealing method to form a sealed edge.

Preferably, the multi-pack is made of thermosynthesized plastic material, preferably ethylene-vinyl acetate copolymer (EVA). EVA has a transparent glossy surface, has good softness and elasticity, has good chemical stability and high temperature resistance, has no biological or chemical activity, is non-toxic, and has good molding processability. It can be autoclaved at 200 ° C and can be transparent O ₂ , CO ₂ , ultraviolet rays, visible light and infrared rays, but impervious to water, with good anti-aging and ozone resistance. It can maintain toughness in liquid nitrogen and is suitable for cryopreservation of cells.

Preferably, the connection port of the sample bag, the connection port of the collection bag, the raw material sampling port, the supplementary sampling port and the liquid replacement filter port are all circular tubular structures;

The axial directions of the connection port of the sample bag, the connection port of the collection bag, the raw material sampling port, the feed feeding port and the liquid replacement filter port are the same.

Preferably, the multi-pack has 1 to 5 sample bags, and the size of the sample bags is 5×5cm;

The multi-pack has 1 to 5 collection bags, and the size of the collection bags is 10×15cm;

Preferably, the multi-pack has 3 sample bags.

Preferably, the multi-pack has 3 collection bags.

Preferably, the connection port of the sample bag, the connection port of the collection bag, the raw material sampling port, the supplementary sampling port and the liquid replacement filter port are respectively equipped with medical sheet clips;

A medical film clip is arranged between the sample bag connecting pipe and the sample bag;

A medical sheet clip is disposed between the collection bag connecting pipe and the collection bag.

Preferably, a medical film clip is disposed between the collection bag and the connection tube of the test sample bag.

The beneficial effects of the utility model are:

1. The body of the culture bag can permeate gas molecules such as oxygen and carbon dioxide, but not water, and at the same time isolate bacteria and other microorganisms to prevent pollution. The gas exchange area is large, which is conducive to a large number of cell cultures in a limited space, and can obtain more How many high-quality cells;

2. The utility model can be applied to cell culture, cell expansion, cell freezing, transgenic, continuous perfusion culture and antibody production, etc., to ensure the aseptic state throughout the process, and effectively reduce the probability of cell contamination and operator infection;

3. The bag body is made of transparent and flexible materials, which is convenient for directly observing the cell state and monitoring the cell culture process under the microscope;

4. It can realize dynamic monitoring of the concentration and viability of cells in the culture bag under different culture time in a closed system. During the cell culture process, sampling, rehydration, and liquid replacement are carried out through heat-sealing machines and aseptic tube machines to ensure that all pipelines are not open, forming a closed culture system that only uses gas molecules such as oxygen and carbon dioxide to prevent contamination by bacteria and microorganisms. Mix the cells in the culture bag body evenly, open the medical clip at the connection port of the sample bag, squeeze in an appropriate amount of cells, close the medical clip, and then heat seal and cut the sample bag connection port of the culture bag body with a heat sealing machine, and the sample bag can be sealed. The cells in the medium are detected without destroying the original closed system;

5. In a closed system, the cultured cells can be collected in different periods of time under different culture times without affecting the continued culture of the remaining cells in the cell culture bag. Mix the cells in the culture bag body, open the medical clip at the connection port of the collection bag, convert the number of cells to be collected in milliliters according to the cell concentration detected in the sample bag, squeeze out the corresponding number of cells, and close the medical clip Clips are used for clinical treatment after heat-sealing and cutting the connection port of the collection bag with a heat-sealing machine. The cells in the collection bag can be used for product quality control, while the remaining cells can continue to be cultured in the culture bag itself. Connect different numbers of collection bags according to needs, generally three are preferred, which is more in line with the cycle of clinical cell therapy. If cytotoxic T cells are used for immunotherapy, the collected cells are transferred to cell culture bags and cultured for 2 weeks, and the cells are collected and reinfused in 3 times, each time 1-3× ¹⁰⁹ CIK cells;

6. The feeding and sampling tube is connected to the syringe to add nutritional factors or cytokines during the cell culture process; the filter tube for changing the liquid is connected to replace the culture medium to realize the liquid change during the culture process.

7. The utility model has the advantages of simple structure, convenient operation, good stability, high reliability, and convenient operation, which is conducive to the cultivation of cells and obtaining more high-quality cells, and can directly perform various operations such as microscopic observation, ensuring The whole process is sterile, and can realize cell expansion, sampling and collection in a closed system, which greatly improves the efficiency of cell culture and reduces the pollution rate, and is suitable for mass expansion of suspension cells and clinical cell therapy.

Description of drawings

Fig. 1 is the structural representation of the utility model;

in:

1-Culture bag body 2-Sample bag system 21-Sample bag connecting tube

22-sample bag 3-collection bag system 31-collection bag connection tube

32-Collection bag 33-Collection bag test sample bag 41-First drop bucket

42-First drop stopper 51-Second drop stopper 52-Second drop stopper

Detailed ways

Below in conjunction with accompanying drawing and specific embodiment, the utility model is described further.

Example 1:

A large-scale cell culture multi-pack as shown in Figure 1, including a culture bag body 1, a sample bag system 2 and a collection bag system 3, one side of the culture bag body 1 is connected to the connection port of the sample bag and the collection bag in sequence Connecting port, raw material sampling port, supplementary sampling port and liquid replacement filter port, of which:

The sample bag system 2 includes a sample bag connecting pipe 21 and a sample bag 22 connected to the sample bag connecting pipe 21, and the sample bag connecting port communicates with the sample bag connecting pipe 21;

The collection bag system 3 includes a collection bag connection pipe 31 and a collection bag 32 connected to the collection bag connection pipe 31 , and the collection bag connection port communicates with the collection bag connection pipe 31 .

The raw material sampling port is connected to the raw material cell bag and the culture bag through a sterile adapter, and the cells to be cultured are transferred into the cell culture bag body 1 .

A first dropping funnel 41 protrudes from the feed feeding port, and a first dropping funnel plug 42 is sleeved on the first dropping funnel 41;

The first dripping funnel 41 and the first dripping funnel plug 42 are detachable structures.

The first dropping funnel 41 on the feeding port is connected with a syringe to add nutritional factors or cytokines to the cell culture bag body 1 .

A second dropping funnel 51 protrudes from the filter port for changing liquid, and a second dropping funnel plug 52 is sleeved on the second dropping funnel 51;

Two layers of filter membranes are sequentially arranged in the second dropping funnel 51 along its axial direction;

The second dropping funnel 51 and the second dropping funnel plug 52 are detachable structures.

The filter membrane in the second dropping funnel 51 close to the second dropping funnel plug 52 is a 70um cell filtration membrane; the filter membrane in the second dropping funnel 51 away from the second dropping funnel plug 52 is a 3um cell filtration membrane.

The second dropping funnel 51 on the filter port for changing the liquid is connected to a negative pressure suction device to realize long-term culture and liquid replacement in the cell culture bag body 1 .

The 70um cell filter membrane is a nylon membrane to prevent large cell clumps from clogging the filter system.

The 3um cell filter membrane is a transwell membrane, which is used to prevent the cultured cells from being sucked away during negative pressure suction.

The collection bag 32 is provided with a collection bag test sample bag 33;

The collection bag 32 is communicated with the collection bag test sample bag 33 through the test sample bag connecting pipe.

The culture bag body 1 is an integral structure.

The culture bag body 1 adopts a heat sealing method to form a sealed edge.

The multi-link bag is made by thermal synthesis of plastic materials, preferably ethylene-vinyl acetate copolymer (EVA). EVA has a transparent glossy surface, has good softness and elasticity, has good chemical stability and high temperature resistance, has no biological or chemical activity, is non-toxic, and has good molding processability. Can be autoclaved at 200°C, permeable to O ₂ , CO ₂ , ultraviolet rays, visible light and infrared rays, but impermeable to water, and has good anti-aging and ozone resistance. It can maintain toughness in liquid nitrogen and is suitable for cryopreservation of cells.

The connection port of the sample bag, the connection port of the collection bag, the sampling port of the raw material, the sampling port of the feeding material and the filter port for changing the liquid are all circular tubular structures;

The axial directions of the connection port of the sample bag, the connection port of the collection bag, the raw material sampling port, the feed feeding port and the liquid replacement filter port are the same.

The multi-pack bag has 1 to 5 sample bags 22, more preferably 3, and the size of the sample bag 22 is 5×5cm;

The multi-pack has 1 to 5 collection bags 32, more preferably 3, and the size of the collection bags 32 is 10×15 cm;

The connecting port of the sample bag, the connecting port of the collection bag, the raw material sampling port, the supplementary sampling port and the liquid replacement filter port are respectively equipped with medical sheet clips;

A medical film clip is arranged between the sample bag connecting pipe 21 and the sample bag 22;

A medical sheet clip is arranged between the collection bag connecting pipe 31 and the collection bag 32 .

A medical sheet clip is disposed between the collection bag 32 and the connecting pipe of the test sample bag.

The multi-pack for large-scale cell culture has the advantages of simple structure, convenient operation, good stability, high reliability, and convenient operation, which is conducive to cell culture and obtaining more high-quality cells, and can directly perform microscopic observation, etc. This kind of operation ensures the whole process of sterility. It can realize cell expansion, sampling and collection in a closed system, which greatly improves the efficiency of cell culture and reduces the pollution rate. It is suitable for mass expansion of suspension cells and clinical cell therapy.

The preferred embodiments of the utility model creation have been specifically described above, but the utility model creation is not limited to the described embodiments, and those skilled in the art can also make Various equivalent modifications or replacements are included within the scope defined by the claims of the present application.

Claims (10)

1. A multi-pack for large-scale cell culture, characterized in that: it comprises a culture bag body (1), a sample bag system (2) and a collection bag system (3), and one side of the culture bag body (1) is sequentially It is connected to the connection port of the sample bag, the connection port of the collection bag, the raw material sampling port, the supplementary sampling port and the liquid replacement filter port, among which: The sample bag system (2) includes a sample bag connecting pipe (21) and a sample bag (22) connected to the sample bag connecting pipe (21), and the sample bag connecting port is connected to the sample bag connecting pipe (21); The collection bag system (3) includes a collection bag connection pipe (31) and a collection bag (32) connected to the collection bag connection pipe (31), and the collection bag connection port communicates with the collection bag connection pipe (31). 2. A kind of large-scale cell culture multiple bag according to claim 1, it is characterized in that: the first dripping funnel (41) protrudes from the described feeding port, and the first dripping funnel (41) is put on Establish the first drop stopper (42); The first dripping funnel (41) and the first dripping funnel plug (42) are detachable structures. 3. A multi-pack for large-scale cell culture according to claim 1, characterized in that: a second dripping funnel (51) protrudes from the filter port for changing liquid, and the second dripping funnel (51) is sleeved The second dropper (52); Two layers of filter membranes are sequentially arranged in the second dropping funnel (51) along its axial direction; The second dropping funnel (51) and the second dropping funnel plug (52) are detachable structures. 4. A kind of large-scale cell culture multi-pack according to claim 3, characterized in that: the filter membrane near the second dropper plug (52) in the second dropper (51) is a 70um cell filter Membrane; the filter membrane far away from the second dropper stopper (52) in the second dropper (51) is a 3um cell filter membrane. 5. a kind of large-scale cell culture multiple bag according to claim 1, is characterized in that: described collection bag (32) is provided with collection bag detection sample bag (33); The collection bag (32) communicates with the collection bag test sample bag (33) through a test sample bag connecting pipe. 6. A multi-pack for large-scale cell culture according to claim 1, characterized in that: said culture bag body (1) has an integrated structure. 7. A multi-pack for large-scale cell culture according to claim 1, characterized in that: the connection port of the sample bag, the connection port of the collection bag, the raw material sampling port, the supplementary sampling port and the filter for changing the liquid The mouths are all circular tubular structures; The axial directions of the connection port of the sample bag, the connection port of the collection bag, the raw material sampling port, the feed feeding port and the liquid replacement filter port are the same. 8. A kind of large-scale cell culture multiple bag according to claim 1, it is characterized in that: described multiple bag has 1～5 sample bag (22), and the size of described sample bag (22) is 5×5cm; The multi-pack has 1 to 5 collection bags (32), and the size of the collection bags (32) is 10×15cm; 9. A large-scale cell culture multi-pack according to claim 1 or 7, characterized in that: the connection port of the sample bag, the connection port of the collection bag, the raw material sampling port, the supplementary sampling port and the replacement port The liquid filter ports are respectively equipped with medical film clips; A medical film clip is arranged between the sample bag connecting pipe (21) and the sample bag (22); A medical sheet clip is arranged between the collection bag connecting pipe (31) and the collection bag (32). 10. A multi-pack for large-scale cell culture according to claim 5, characterized in that: a medical film clip is arranged between the collection bag (32) and the connecting tube of the test sample bag.