Cell culture bag

Abstract

The invention discloses a cell culture bag. The cell culture bag comprises a plastic bag body and at least two joints arranged on the plastic bag body. The connector can selectively communicate the culture space with the outside through fluid. The plastic bag body is made of a double-layer film, the double-layer film is made of polyolefin and comprises a first layer body and a second layer body, and the melting point of the first layer body is higher than that of the second layer body. The plastic bag body comprises a culture space and a breathable surface. The culture space is formed inside the plastic bag body, and the second layer body faces the culture space. The air-permeable surface is communicated with the culture space and the outside through air, and the culture space per unit volume is provided with the air-permeable surface of 0.6-1.5 cm < 2 > / ml. The oxygen penetration rate of the cell culture bag is greater than 200g / m < 2 > / day and the water vapor penetration rate is less than 10g / m < 2 > / day under the environment of 37 DEG C and 1 atmospheric pressure.

Description

Technical Field

[0001] The present invention relates to a cell culture bag, and more particularly to a cell culture bag with good air permeability. Background Art

[0002] When performing cell therapy, cells taken out from a patient's body must be placed in a cell culture Erlenmeyer flask for cultivation. After the cells grow to a certain number, they are taken out and injected into the human body.

[0003] During the growth process, cells need oxygen and also emit carbon dioxide. Therefore, in order to provide an appropriate growth environment for the cells, the cell culture Erlenmeyer flask should not be filled with liquid to supply the gas required for cell growth. And the liquid level in the cell culture Erlenmeyer flask should not be higher than the bottle mouth to prevent the liquid from flowing out of the bottle mouth.

[0004] In order to achieve the effect of gas exchange, generally, an air-permeable membrane is attached to the bottle mouth of the cell culture Erlenmeyer flask and placed in an incubator. By adjusting the gas environment in the incubator, gas can pass through the air-permeable membrane and enter and exit the cell culture Erlenmeyer flask to achieve the effect of controlling the cell growth environment.

[0005] Due to the limitations of the above structure, the volume of a general cell culture Erlenmeyer flask is relatively large, resulting in inconvenient use. Therefore, how to overcome the defect of the large volume of the cell culture Erlenmeyer flask through the improvement of the structural design has become one of the important issues to be solved in this field. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to provide a cell culture bag in view of the deficiencies of the prior art.

[0007] To solve the above technical problem, one of the technical solutions adopted by the present invention is to provide a cell culture bag. The cell culture bag includes a plastic bag body and at least two connectors provided on the plastic bag body. The connectors can selectively fluid-connect the culture space with the outside. The plastic bag body is made of a double-layer film. The material of the double-layer film includes polyolefin. The double-layer film includes a first layer body and a second layer body. The melting point of the first layer body is higher than that of the second layer body. The plastic bag body includes a culture space and a breathable surface. The culture space is formed inside the plastic bag body, and the second layer body faces the culture space. The breathable surface is gas-connected to the culture space and the outside. Each unit volume of the culture space has a breathable surface of 0.6 square centimeters per milliliter to 1.5 square centimeters per milliliter. The oxygen penetration rate of the cell culture bag under the environment of 37 °C and 1 atmosphere is greater than 200 g / m 2 / day, and the water vapor penetration rate is less than 10 g / m 2 / day.

[0008] Furthermore, the thickness of the double-layer film is 50 micrometers to 140 micrometers.

[0009] Further, the thickness ratio of the first layer body to the second layer body is 6 to 12.

[0010] Further, the plastic bag body further includes a sealed edge section, which is formed by heat-sealing the second layer body. The sealed edge section defines the range of the culture space and surrounds the breathable surface.

[0011] Further, the width of the sealed edge section is 6 mm to 15 mm.

[0012] Further, the material of the first layer body is linear low density polyethylene with a melting point of 116°C to 125°C.

[0013] Further, the weight average molecular weight of the linear low density polyethylene is 132,000 g / mol to 134,000 g / mol.

[0014] Further, the material of the second layer body is low density polyethylene with a melting point of 106°C to 115°C.

[0015] Further, the weight average molecular weight of the low density polyethylene is 129,000 g / mol to 131,000 g / mol.

[0016] Further, the culture space is used to hold a culture liquid, and the culture liquid has an average breathable surface of 1.0 square centimeters per milliliter to 1.9 square centimeters per milliliter per unit volume.

[0017] Further, the total volume of the culture space is 1000 milliliters to 2000 milliliters, and the total volume of the culture liquid is 700 milliliters to 1200 milliliters.

[0018] To solve the above technical problems, another technical solution adopted by the present invention is to provide a cell culture bag. The cell culture bag includes a plastic bag body and at least two connectors provided on the plastic bag body. The connectors can selectively fluid-connect the culture space with the outside. The material of the plastic bag body is selected from the group consisting of ethylene vinyl acetate copolymer, linear low density polyethylene, and low density polyethylene. The plastic bag body has a culture space and a breathable surface. The culture space is formed inside the plastic bag body, and the breathable surface is gas-connected to the culture space and the outside. Each unit volume of the culture space has a breathable surface of 0.6 square centimeters per milliliter to 1.5 square centimeters per milliliter. The oxygen permeability of the cell culture bag at 37°C and 1 atmosphere is greater than 200 g / m 2 / day, and the water vapor permeability is less than 10 g / m 2 / day.

[0019] Further, the material of the plastic bag body is ethylene vinyl acetate copolymer with a melting point of 80°C to 90°C.

[0020] Furthermore, the weight-average molecular weight of the ethylene-vinyl acetate copolymer is from 31,000 g / mol to 33,000 g / mol.

[0021] One beneficial effect of the present invention is that the cell culture bag provided by the present invention can, through the technical solutions of "material selection of the plastic bag body" and "having a gas-permeable surface of 0.6 square centimeters per milliliter to 1.5 square centimeters per milliliter for each unit volume of the culture space", enable the cell culture bag to have good oxygen permeability and appropriate water vapor permeability.

[0022] To further understand the features and technical content of the present invention, please refer to the following detailed description and drawings of the present invention. However, the provided drawings are only for reference and illustration, and are not used to limit the present invention. Description of the Drawings

[0023] Figure 1 It is a side view schematic diagram of the cell culture bag of the present invention.

[0024] Figure 2 For the cell culture bag of one embodiment of the present invention in Figure 1 the cross-sectional schematic diagram of the II-II section.

[0025] Figure 3 For the cell culture bag of another embodiment of the present invention in Figure 1 the cross-sectional schematic diagram of the II-II section. Detailed Embodiments

[0026] The following are specific examples to illustrate the embodiments of the "cell culture bag" disclosed by the present invention. Those skilled in the art can understand the advantages and effects of the present invention from the content disclosed in this specification. The present invention can be implemented or applied through other different specific embodiments, and various details in this specification can also be modified and changed based on different viewpoints and applications without departing from the concept of the present invention. Additionally, the drawings of the present invention are only simple schematic illustrations and are not drawn according to actual dimensions, hereby stated in advance. The following embodiments will further detail the related technical content of the present invention, but the disclosed content is not used to limit the protection scope of the present invention. Additionally, the term "or" used herein should, depending on the actual situation, possibly include any one or a combination of more of the related listed items.

[0027] In order to overcome the problem that the volume of the conventional cell culture Erlenmeyer flask is too large, the present invention provides a cell culture bag, which has good air permeability and can directly exchange gases with the outside world. Therefore, the cell culture bag of the present invention can overcome the problem that the conventional cell culture Erlenmeyer flask has a large volume to accommodate air, and has the effect of being convenient to use.

[0028] Please refer toFigure 1 As shown, the cell culture bag of the present invention has a plastic bag body 1 and at least two connectors. The connectors are arranged on the plastic bag body 1 to facilitate the injection of culture medium, sampling, or the removal of culture medium. The types of connectors can be selected according to the usage purpose. For the sake of convenience of explanation, Figure 1 the connectors in

[0029] can include a feeding connector 2, a sampling connector 3, and a discharging connector 4.

[0030] Please refer jointly to Figure 1 and Figure 2 As shown, the plastic bag body 1 is a transparent bag body to observe the cell state in the culture space 10. Specifically, the light transmittance of the plastic bag body 1 is above 90%, and the haze of the plastic bag body 1 is less than 11%.

[0031] A culture space 10 is formed inside the plastic bag body 1. The culture space 10 can be used to hold a culture liquid to help cells grow in a sterile environment. The setting of the connectors can selectively connect the culture space 10 with the outside fluid to facilitate the injection of the culture liquid, sampling, or the removal of the culture liquid.

[0032] In order to form the culture space 10 inside the plastic bag body 1, first take a film material and perform a heat-sealing procedure after folding. The parts of the film material processed by the heat-sealing procedure will be combined due to high-temperature melting, and a sealing section 11 on the plastic bag body 1 will be formed.

[0033] Except for the positions where the connectors are set, the sealing section 11 will form a closed area to define the range of the culture space 10. When the range formed by the closure of the sealing section 11 is larger, the culture space 10 formed inside the plastic bag body 1 is larger; when the range formed by the closure of the sealing section 11 is smaller, the culture space 10 formed inside the plastic bag body 1 is smaller. In an exemplary embodiment, the volume of the culture space 10 is 1000 milliliters to 2000 milliliters, but the present invention is not limited thereto. In an exemplary embodiment, the width W of the sealing section 11 is 6 millimeters to 15 millimeters, but the present invention is not limited to this.

[0034] Through holes can be formed on the plastic bag body 1 so that the cell culture bag can be used in a hanging state. When used in a hanging state, in order to prevent the culture liquid from leaking out of the through holes, a leak-proof section 12 can also be formed on the plastic bag body 1. In an exemplary embodiment, the leak-proof section 12 is integrally formed with the sealing section 11 and jointly surrounds the through holes. Therefore, the through holes will not be connected to the culture space 10.

[0035] In other embodiments, the leak-proof section 12 may also be independently formed on the plastic bag body 1 and surround the through hole alone, but the present invention is not limited thereto.

[0036] During the process of cell growth, oxygen is required for respiration and carbon dioxide is also emitted. Therefore, the plastic bag body 1 needs to be made of a breathable material. Specifically, both sides of the plastic bag body 10 respectively have a breathable surface S, and the breathable surface S is surrounded by the sealing section 11. The breathable surfaces S are respectively located on both sides of the culture space 10. In this way, the culture space 10 can communicate with the external gas through the breathable surfaces S on both sides to achieve the effect of gas exchange.

[0037] In order to avoid the problem that cells die due to insufficient oxygen caused by too low air exchange rate during cell culture, the present invention controls that the culture space 10 has an average of 0.6 square centimeters per milliliter to 1.5 square centimeters per milliliter of breathable surface S per unit volume. For example: the culture space 10 has an average of 0.8 square centimeters per milliliter, 1.0 square centimeters per milliliter, 1.2 square centimeters per milliliter or 1.4 square centimeters per milliliter of breathable surface S per unit volume. Preferably, the culture space 10 has an average of 0.65 square centimeters per milliliter to 1.35 square centimeters per milliliter of breathable surface S per unit volume.

[0038] For example, in a demonstration embodiment, the total area of the breathable surfaces S on both sides of the plastic bag body 10 is 1314 square centimeters. When the culture space 10 is 1000 milliliters, the culture space 10 has an average of 1.314 square centimeters per milliliter of breathable surface S per unit volume. When the culture space 10 is 2000 milliliters, the culture space 10 has an average of 0.657 square centimeters per milliliter of breathable surface S per unit volume.

[0039] For the convenience of use and to improve the survival rate of cells, during use, the culture liquid does not completely fill the culture space 10. For example, the added amount of the culture liquid can be 50% to 75% of the total volume of the culture space 10. In a demonstration embodiment, the added volume of the culture liquid is 700 milliliters to 1200 milliliters, but the present invention is not limited thereto.

[0040] After experimental tests, when the culture liquid has an average of 0.9 square centimeters per milliliter to 1.9 square centimeters per milliliter of breathable surface S per unit volume, a balance can be achieved between the cell survival rate and the size of the cell culture bag. For example: the culture liquid has an average of 1.0 square centimeters per milliliter, 1.2 square centimeters per milliliter, 1.4 square centimeters per milliliter, 1.6 square centimeters per milliliter or 1.8 square centimeters per milliliter of breathable surface S per unit volume.

[0041] For example, in a demonstration embodiment, the total area of the breathable surfaces S on both sides of the plastic bag body 10 is 1314 square centimeters. When the addition amount of the culture liquid is 700 milliliters, the culture liquid per unit volume has an average breathable surface S of 1.877 square centimeters per milliliter. When the addition amount of the culture liquid is 1200 milliliters, the culture liquid per unit volume has an average breathable surface S of 1.095 square centimeters per milliliter.

[0042] To improve the breathability of the plastic bag body 1, the material of the plastic bag body 1 may include at least one of ethylene-vinyl acetate copolymer (EVA), linear low density polyethylene (LLDPE), or low density polyethylene (LDPE).

[0043] Specifically, the melting point of the ethylene-vinyl acetate copolymer measured by differential scanning calorimetry (DSC) is 80°C to 90°C. Preferably, the melting point of the ethylene-vinyl acetate copolymer is 82°C to 86°C. The weight average molecular weight of the ethylene-vinyl acetate copolymer measured by gel permeation chromatography (GPC) is 31,000 grams per mole to 33,000 grams per mole. Preferably, the weight average molecular weight of the ethylene-vinyl acetate copolymer is 31,500 grams per mole to 32,500 grams per mole. The melt index of the ethylene-vinyl acetate copolymer measured according to the ASTM D1238 standard test method is 1.6 to 2.0. Preferably, the melt index of the ethylene-vinyl acetate copolymer is 1.7 to 1.9. The density of the ethylene-vinyl acetate copolymer measured according to the ASTM D1505 standard test method is 0.910 grams per cubic centimeter to 0.950 grams per cubic centimeter. Preferably, the density of the ethylene-vinyl acetate copolymer is 0.935 grams per cubic centimeter to 0.940 grams per cubic centimeter.

[0044] Specifically, the melting point of linear low density polyethylene measured by differential scanning calorimetry is 116°C to 125°C. Preferably, the melting point of linear low density polyethylene is 120°C to 124°C. The weight average molecular weight of linear low density polyethylene measured by gel permeation chromatography is 132,000 g / mol to 134,000 g / mol. Preferably, the weight average molecular weight of linear low density polyethylene is 132,500 g / mol to 133,500 g / mol. The melt index of linear low density polyethylene measured according to the ASTM D1238 standard test method is 1.8 to 2.2. Preferably, the melt index of linear low density polyethylene is 1.9 to 2.1. The density of linear low density polyethylene measured according to the ASTM D1505 standard test method is 0.910 g / cm³ to 0.950 g / cm³. Preferably, the density of linear low density polyethylene is 0.915 g / cm³ to 0.925 g / cm³.

[0045] Specifically, the melting point of low density polyethylene measured by differential scanning calorimetry is 106°C to 115°C. Preferably, the melting point of low density polyethylene is 109°C to 113°C. The weight average molecular weight of low density polyethylene measured by gel permeation chromatography is 129,000 g / mol to 131,000 g / mol. Preferably, the weight average molecular weight of low density polyethylene is 129,500 g / mol to 130,500 g / mol. The melt index of low density polyethylene measured according to the ASTM D1238 standard test method is 1.8 to 2.2. Preferably, the melt index of low density polyethylene is 1.9 to 2.1. The density of low density polyethylene according to the ASTM D1505 standard test method is 0.910 g / cm³ to 0.950 g / cm³. Preferably, the density of low density polyethylene is 0.920 g / cm³ to 0.925 g / cm³.

[0046] Please refer to Figure 2 As shown, the thickness T of the plastic bag body 1 also affects the air permeability. When the thickness T of the plastic bag body 1 is 50 microns to 140 microns, the plastic bag body 1 can have good air permeability. Specific experimental descriptions will be described later.

[0047] Through the selection of materials and the adjustment of thickness, the oxygen permeability of the plastic bag body 1 at 37°C and 1 atmospheric pressure can be greater than 200 g / m² / day, and the water vapor permeability of the plastic bag body 1 at 37°C and 1 atmospheric pressure can be less than 10 g / m² / day.

[0048] In order to confirm that the cell culture bag of the present invention has good air permeability, ethylene vinyl acetate copolymer, linear low density polyethylene and low density polyethylene are respectively used to make plastic bag bodies with different thicknesses (Test Examples 1 to 9), and the plastic bag bodies are subjected to characteristic tests. The test results are listed in Table 1.

[0049] In Table 1, the water vapor transmission rate of the plastic bag body was measured according to the ASTM F1249 standard test method under the environment of 37°C and 1 atmosphere. The oxygen transmission rate of the plastic bag body was measured according to the ASTM D3985 standard test method under the environment of 37°C and 1 atmosphere. In addition, a haze meter (ASTM D1003) was used to measure the light transmittance and haze of the plastic bag body, a surface resistance meter was used to measure the surface impedance of the plastic bag body, and a universal tensile testing machine (ASTM D882) was used to measure the tensile strength of the plastic bag body.

[0050] Table 1

[0051]

[0052] From the results in Table 1, it can be known that when the thickness of the plastic bag body is relatively thick, the oxygen transmission rate of the plastic bag body will decrease and cannot meet the expected oxygen transmission rate (greater than 200 g / m² / day). When the thickness of the plastic bag body is relatively thin, although the oxygen transmission rate of the plastic bag body is sufficient to supply cell growth, the water vapor transmission rate of the plastic bag body will also increase. When the evaporation rate of water vapor is too fast, the concentration of the culture liquid is too high, which is likely to cause cell death. Therefore, under the comprehensive evaluation of the oxygen transmission rate and the water vapor transmission rate, the thickness of the plastic bag body is preferably 50 to 140 microns.

[0053] From the results in Table 1, it can be known that selecting ethylene-vinyl acetate copolymer, linear low-density polyethylene or low-density polyethylene as the material of the plastic bag body can not only make the plastic bag body have appropriate oxygen transmission rate and water vapor transmission rate, but also make the plastic bag body have appropriate light transmittance (greater than 90%) and haze (less than 11%).

[0054] Moreover, the tensile strength of the plastic bag body can be greater than 3 kgf. Preferably, the plastic bag body can have a tensile strength of 3 to 5.5 kgf. In this way, it can be avoided that the plastic bag body is broken due to external force during processing operations. Since the impedance value of the cell culture bag is generally measured to infer the cell growth status, the surface impedance of the plastic bag body needs to be greater than 2×10 12 , so as to avoid the plastic bag body affecting the measurement results.

[0055] The present invention provides a cell culture bag of another embodiment, which has a structure similar to that of the Figure 1 cell culture bag. The cell culture bag has a plastic bag body 1, and a feeding joint 2, a sampling joint 3 and a discharging joint 4 arranged on the plastic bag body 1. The difference is that the plastic bag body 1 is made of a double-layer film.

[0056] Please refer to Figure 3As shown, after the double-layer film is processed by the heat-sealing process, the part of the double-layer film processed by the heat-sealing process will be combined due to high-temperature melting, and a sealing section 11 on the plastic bag body 1 will be formed. The sealing section 11 defines the range of the culture space 10 and surrounds the breathable surface S. In an exemplary embodiment, the thickness of the double-layer film is 50 to 140 micrometers.

[0057] Specifically, the double-layer film includes a first layer body 101 and a second layer body 102, and the first layer body 101 is disposed on the second layer body 102. The second layer body 102 is located inside the plastic bag body 1, that is to say, the second layer body 102 faces the culture space 10. Therefore, in the heat-sealing process, the second layer body 102 is the main part that is melted and joined to form the sealing section 11.

[0058] In order to achieve an ideal breathable effect, the thickness T of the double-layer film cannot be greater than 150 micrometers. Therefore, in the heat-sealing process, the double-layer film is prone to the problem of film breakage. To solve this problem, the present invention combines film layers with different melting points to manufacture the double-layer film. Among them, the melting point of the first layer body 101 is higher than that of the second layer body 102. In this way, a lower heat-sealing temperature can be used in the heat-sealing process to form the sealing section 11, thereby avoiding the occurrence of film breakage.

[0059] Due to the difference in melting points, the double-layer film can have good breathability and a lower processing temperature. In an exemplary embodiment, the material of the first layer body 101 is linear low-density polyethylene, and the second layer body 102 is low-density polyethylene. The materials of the first layer body 101 and the second layer body 102 are similar in type, so the double-layer film can be made by co-extrusion. The main difference between the first layer body 101 and the second layer body 102 is the different melting points. The melting point of the first layer body 101 can be 3°C to 20°C higher than that of the second layer body 102. Preferably, the melting point of the first layer body 101 can be 5°C to 15°C higher than that of the second layer body 102.

[0060] Through experimental tests, when the ratio of the thickness of the first layer body 101 to the thickness of the second layer body 102 is 6 to 12, the double-layer film can take into account both breathability and good heat-sealing effect. Preferably, the ratio of the thickness of the first layer body 101 to the thickness of the second layer body 102 is 8 to 10.

[0061] In order to confirm that the cell culture bag of the present invention has good breathability, linear low-density polyethylene and low-density polyethylene are first made into a 100-micrometer double-layer film by co-extrusion, and then the double-layer film is used to make a plastic bag body (Test Examples 10 to 12). The double-layer films in Test Examples 10 to 12 have different thickness ratios of the first layer body 101 and the second layer body 102.

[0062] In addition, the water vapor transmission rate, oxygen transmission rate, light transmittance, haze, surface impedance, tensile strength, and heat-sealing bag breakage rate of the plastic bag body were tested, and the test results are listed in Table 2. Among them, the test methods for the water vapor transmission rate, oxygen transmission rate, light transmittance, haze, surface impedance, and tensile strength are as described above and will not be elaborated here.

[0063] In the test of the heat-sealing bag breakage rate, the plastic bag body was heat-sealed at a temperature of 115 °C to make a cell culture bag. 70% of the liquid was filled in the cell culture bag, and a weight of 1 kg was placed on the cell culture bag to observe whether the liquid leaked out of the cell culture bag. The above-mentioned tests of filling the liquid and placing the weight were carried out on multiple plastic bag bodies respectively, and the number of times was counted to represent the bag breakage rate of the cell culture bag as a proportion. For example, a bag breakage rate of 15% means that in twenty tests, the cell culture bag ruptured three times.

[0064] Table 2

[0065]

[0066] From the results in Table 2, it can be seen that when the thickness ratio of the first layer body to the second layer body is 9:1, the cell culture bag has the lowest heat-sealing bag breakage rate. Moreover, the heat-sealing bag breakage rate of the cell culture bag increases as the thickness ratio of the second layer body increases.

[0067] From the results in Table 1 and Table 2, it can be seen that by setting the first layer body and the second layer body, the cell culture bag can have a better heat-sealing effect and is not prone to bag breakage while maintaining the expected oxygen transmission rate and water vapor transmission rate.

[0068] [Advantages of the Embodiment]

[0069] One of the advantages of the present invention is that the cell culture bag provided by the present invention can have a good oxygen transmission rate and an appropriate water vapor transmission rate through the technical solutions of "material selection of the plastic bag body" and "having an air-permeable surface of 0.6 square centimeters per milliliter to 1.5 square centimeters per milliliter for each unit volume of the culture space".

[0070] Furthermore, in order to improve the manufacturing yield of the cell culture bag while maintaining appropriate air permeability, a double-layer film can be used to manufacture the cell culture bag. By laminating two layer bodies with different melting points, the processing temperature in the heat-sealing process can be reduced, and the double-layer film can be prevented from breaking during the heat-sealing process.

[0071] The content disclosed above is only the preferred feasible embodiment of the present invention, and does not limit the protection scope of the claims of the present invention. Therefore, all equivalent technical changes made by using the content of the specification and drawings of the present invention are included in the protection scope of the claims of the present invention.

Claims

1. A cell culture bag, characterized in that: The cell culture bag comprises: A plastic bag, whose material includes polyolefin, is made of a double-layer film, the double-layer film includes a first layer and a second layer, the melting point of the first layer is higher than the melting point of the second layer, and the plastic bag includes: a culture space formed inside the plastic bag body, wherein the second layer faces the culture space; and a gas-permeable surface, the gas of which connects the culture space with the outside world; The culture space has an average gas permeable surface area of ​​0.6 cm2 / ml to 1.9 cm2 / ml per unit volume; and At least two connectors are provided on the plastic bag body, and the connectors can selectively connect the culture space with the outside through fluid; The oxygen permeability of the cell culture bag under the conditions of 37° C. and 1 atmosphere pressure is greater than 200 g / m 2 / day, and the water vapor permeability is less than 10 g / m 2 / day.

2. The cell culture bag according to claim 1, characterized in that: The thickness of the double-layer film is 50 micrometers to 140 micrometers.

3. The cell culture bag according to claim 1, characterized in that: The thickness ratio of the first layer to the second layer is 6 to 12.

4. The cell culture bag according to claim 1, characterized in that: The plastic bag body further comprises a sealing edge section, which is formed by heat-sealing the second layer. The sealing edge section defines the range of the culture space and surrounds the air-permeable surface.

5. The cell culture bag according to claim 4, characterized in that: The width of the edge sealing section is 6 mm to 15 mm.

6. The cell culture bag according to claim 1, characterized in that: The material of the first layer is linear low-density polyethylene with a melting point of 116° C. to 125° C.

7. The cell culture bag according to claim 6, characterized in that: The linear low density polyethylene has a weight average molecular weight of 132,000 to 134,000 g / mol.

8. The cell culture bag according to claim 1, characterized in that: The material of the second layer is low-density polyethylene with a melting point of 106°C to 115°C.

9. The cell culture bag according to claim 8, characterized in that: The low density polyethylene has a weight average molecular weight of 129,000 g / mol to 131,000 g / mol.

10. The cell culture bag according to claim 1, characterized in that: The culture space is used to contain a culture liquid, and the culture liquid has an average gas permeable surface of 1.0 cm2 / ml to 1.9 cm2 / ml per unit volume.

11. The cell culture bag according to claim 10, characterized in that: The total volume of the culture space is 1,000 ml to 2,000 ml, and the total volume of the culture liquid is 700 ml to 1,200 ml.

12. A cell culture bag, characterized in that: The cell culture bag comprises: A plastic bag, the material of which is selected from the group consisting of ethylene vinyl acetate copolymer, linear low-density polyethylene and low-density polyethylene, the plastic bag comprising: a culture space formed inside the plastic bag; and a gas-permeable surface, the gas of which connects the culture space with the outside world; wherein the culture space has a gas permeable surface area of ​​0.6 cm2 / ml to 1.5 cm2 / ml per unit volume; and At least two connectors are provided on the plastic bag body, and the connectors can selectively connect the culture space with the outside through fluid; The oxygen permeability of the cell culture bag under 37°C and 1 atmosphere pressure is greater than 200 g / m2 / day, and the water vapor permeability is less than 10 g / m 2 / day.

13. The cell culture bag according to claim 12, characterized in that: The material of the plastic bag body is ethylene vinyl acetate copolymer with a melting point of 80° C. to 90° C.

14. The cell culture bag according to claim 13, characterized in that: The ethylene vinyl acetate copolymer has a weight average molecular weight of 31,000 g / mol to 33,000 g / mol.

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