Cell culture bag

Abstract

To provide a cell culture bag.SOLUTION: A cell culture bag includes a plastic bag body and at least two connectors disposed on the plastic bag body. The connectors may provide liquid communication between a culture space and an external environment. The cell culture bag is made of a bilayer film using polyolefin, the bilayer film including a first layer and a second layer, and the melting point of the first layer is higher than the melting point of the second layer. The plastic bag body includes a culture space and a breathable surface. The culture space is formed inside the plastic bag, and the second layer faces the culture space. The culture space and the external environment are gas-communicated through the breathable surface, and the average area of the breathable surface per unit volume of the culture space is 0.6 cm2 / mL to 1.5 cm2 / mL. The oxygen transmission rate of the cell culture bag in an environment of 37°C and 1 atm is more than 200 g / m2 / day, and its water vapor transmission rate is less than 10 g / m2 / day.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a cell culture bag, and particularly to a cell culture bag having good air permeability.

Background Art

[0002] During cell therapy, cells taken from a patient's body need to be cultured in a cell culture flask. After the cells have grown to a certain number, they are taken out and injected into the human body.

[0003] In the process of cell growth, cells require oxygen and also emit carbon dioxide. Therefore, in order to provide an appropriate growth environment for cells, the cell culture flask should not be completely filled with liquid so as to provide the gases necessary for cell growth. Also, the liquid level in the cell culture flask should not be higher than the mouth of the bottle to prevent the liquid from flowing out of the mouth of the bottle.

[0004] In order to achieve the effect of gas exchange, the cell culture flask has a breathable membrane attached to its mouth and is placed in an incubator. By adjusting the gas environment inside the incubator, the gas can enter the cell culture flask through the breathable film so as to achieve the effect of controlling the cell growth environment.

[0005] Due to the limitations of the above structure, the volume of the cell culture flask is often large and it becomes difficult to use. Therefore, overcoming the drawback of the large volume of the cell culture flask by improving the structural design is an important issue to be solved in this industry.

Summary of the Invention

Problems to be Solved by the Invention

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

Means for Solving the Problems

[0007] To solve the above technical problems, one technical means adopted by the present invention is to provide a cell culture bag including a plastic bag body including a culture space and a breathable surface, and at least two connecting parts installed on the plastic bag body. The culture space and the external environment can be in liquid communication through the connecting parts. The plastic bag body is made of a two-layer film including polyolefin. The two-layer film includes a first layer and a second layer. The melting point of the first layer is higher than that of the second layer. The culture space is formed inside the plastic bag. The second layer faces the culture space. The culture space and the external environment are in gas communication through the breathable surface. The average area of the breathable surface per unit volume of the culture space is 0.6 cm 2 / mL to 1.5 cm 2 / mL. The oxygen permeability of the cell culture bag at 37 °C and 1 atm is more than 200 g / m 2 / day, and the water vapor permeability is less than 10 g / m 2 / day.

[0008] In one embodiment, the thickness of the two-layer film is 50 μm to 140 μm.

[0009] In one embodiment, the value of the thickness ratio of the first layer to the second layer (the first layer / the second layer) is 6 to 12.

[0010] In one embodiment, the plastic bag includes a sheet part formed by heat-sealing to the second layer, and the range of the culture space is defined by the sheet part so as to surround the breathable surface.

[0011] In one embodiment, the width of the sheet part is 6 mm to 15 mm.

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

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

[0014] In one embodiment, the material of the second layer is low-density polyethylene with a melting point of 106°C to 115°C.

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

[0016] In one embodiment, the culture space contains a culture solution, and the average area of the breathable surface per unit volume of the culture solution is 1.0 cm 2 / mL to 1.9 cm 2 / mL.

[0017] In one embodiment, the total volume of the culture space is 1000 mL to 2000 mL, and the total volume of the culture solution is 700 mL to 1200 mL.

[0018] To solve the above technical problems, another technical means adopted by the present invention is to provide a cell culture bag.

[0019] To solve the above technical problems, another technical means adopted by the present invention is to provide a cell culture bag including a plastic bag body including a culture space and a breathable surface, and at least two connection parts installed on the plastic bag body. The culture space and the external environment can be in liquid communication through the connection parts, the material of the plastic bag is selected from the group consisting of ethylene vinyl acetate copolymer, linear low-density polyethylene, and low-density polyethylene, the culture space is formed inside the plastic bag, the culture space and the external environment are in gas communication through the breathable surface, the average area of the breathable surface per unit volume of the culture space is 0.6 cm 2 / mL to 1.5 cm 2 / mL, and the oxygen permeability of the cell culture bag at 37°C and 1 atm is more than 200 g / m 2 / day, and the water vapor permeability is less than 10 g / m 2 / day.

[0020] In one embodiment, the material of the plastic bag is an ethylene-vinyl acetate copolymer with a melting point of 80°C to 90°C.

[0021] In one embodiment, the weight average molecular weight of the ethylene-vinyl acetate copolymer is 31,000 g / mol to 33,000 g / mol.

Advantages of the Invention

[0022] One of the advantageous effects of the present invention is that the cell culture bag according to the present invention has good oxygen permeability and appropriate water vapor permeability due to technical features such as "selection of the material of the plastic bag" and "the average area of the breathable surface per unit volume of the culture space is 0.6 cm 2 / mL to 1.5 cm 2 / mL".

Brief Description of the Drawings

[0023]

Figure 1

Figure 2

Figure 3

Modes for Carrying Out the Invention

[0024] For a better understanding of the features and technical content of the present invention, please refer to the following detailed description of the present invention and the accompanying drawings. However, the provided accompanying drawings are only for reference and explanation, and are not for limiting the scope of the claims of the present invention.

[0025] The following will describe the "cell culture bag" according to the present invention by means of specific embodiments, and those skilled in the art can understand the advantages and effects of the present invention based on the content disclosed in this specification. The present invention can be implemented or applied by other different specific embodiments, and for each detail in this specification, various modifications and changes can be made based on different viewpoints and applications without departing from the concept of the present invention. Also, as explained in advance, the attached drawings of the present invention are simple schematic explanations and are not drawn based on actual sizes. The technical content of the present invention will be further described in detail based on the following embodiments, but the disclosed content does not limit the protection scope of the present invention. Also, the term "or" used in this specification may include any one or a combination of multiple items listed in relation according to the actual situation. In order to overcome the problem that the volume of the conventional cell culture flask is too large, the cell culture bag according to the present invention has good air permeability and can directly exchange gas with the external environment. Therefore, since the cell culture bag according to the present invention can accommodate air, the problem of its large volume can be overcome and it becomes easier to use.

[0026] As shown in FIG. 1, the cell culture bag according to the present invention includes a plastic bag 1 and at least two connecting parts. The connecting parts are installed on the plastic bag 1 to facilitate the injection, sampling, or extraction of the culture solution. The type of the connecting part (connector) can be selected according to the purpose. For the convenience of explanation, the connecting parts in FIG. 1 may include a feeding connecting part 2, a sampling connecting part 3, and a discharging connecting part 4.

[0027] For example, the feeding connecting part 2 and the discharging connecting part 4 can adopt needleless connectors, and the sampling connecting part 3 can adopt polybase male luer connectors, but the present invention is not limited thereto. When actually applied, in order to avoid contamination, it is necessary to cut off the connecting part that has been used once.

[0028] As shown in FIGS. 1 and 2, the plastic bag 1 is a transparent bag so as to observe the cell state inside the culture space 10. Specifically, the light transmittance of the plastic bag 1 is 90% or more, and the haze value of the plastic bag 1 is less than 11%.

[0029] Inside the plastic bag 1, a culture space 10 for accommodating a culture solution is formed so that cells can grow in a sterile environment. By installing a connection part, the culture space 10 and the external environment are communicated, which facilitates the injection, sampling or removal of the culture solution.

[0030] In order to form the culture space 10 inside the plastic bag 1, after folding the film material, a heat sealing process is performed. The parts of the film material processed by the heat sealing process are joined to each other by high-temperature melting to form the sheet part 11 of the plastic bag 1.

[0031] In addition to setting the position of the connection part, a closed area is formed by the sheet part 11 so as to define the range of the culture space 10. The larger the closed area formed by the sheet part 11, the larger the culture space 10 formed inside the plastic bag 1. On the other hand, the smaller the closed area formed by the sheet part 11, the smaller the culture space 10 formed inside the plastic bag 1. In one exemplary embodiment, the volume of the culture space 10 is 1000 mL to 2000 mL, but the present invention is not limited thereto. In one exemplary embodiment, the width W of the sheet part 11 is 6 mm to 15 mm, but the present invention is not limited thereto.

[0032] A through hole may be formed in the plastic bag 1 so that the cell culture bag can be used in a suspended state. When used in a suspended state, a leak prevention part 12 may be further formed in the plastic bag 1 so as to avoid the culture solution oozing out from the through hole. In one exemplary embodiment, the leak prevention part 12 and the sheet part 11 are integrally formed so as to surround the through hole. Therefore, the through hole does not communicate with the culture space 10.

[0033] In other embodiments, the leak prevention portion 12 may be independently formed on the plastic bag 1 so as to surround the through hole by itself, but the present invention is not limited thereto.

[0034] In the process of cell growth, cells may inhale oxygen and exhale carbon dioxide. Therefore, the plastic bag 1 needs to be made of a breathable material. Specifically, breathable surfaces S are respectively formed on both sides of the plastic bag 10. Thereby, the culture space 10 can achieve the effect of gas exchange by communicating with the external gas through the breathable surfaces S on both sides.

[0035] In order to avoid the problem that the ventilation rate is too low during the cell culture process and the cells die due to oxygen deficiency, in the present invention, the average area of the breathable surface S per unit volume of the culture space 10 is 0.6 cm 3 / mL to 1.5 cm 3 / mL. The average area of the breathable surface S per unit volume of the culture space 10 is 0.8 cm 3 / mL, 1.0 cm 3 / mL, 1.2 cm 3 / mL or 1.4 cm 3 / mL. Preferably, the average area of the breathable surface S per unit volume of the culture space 10 is 0.65 cm 3 / mL to 1.35 cm 3 / mL.

[0036] For example, in one exemplary case, the total area of the breathable surfaces S on both sides of the plastic bag 1 is 1314 cm 2 . When the culture space 10 is 1000 mL, the average area of the breathable surface S per unit volume of the culture space 10 is 1.314 cm 2 / mL. When the culture space 10 is 2000 mL, the average area of the breathable surface S per unit volume of the culture space 10 is 0.657 cm 2 / mL.

[0037] To improve ease of use and cell viability, during use, the culture medium does not completely fill the culture space 10. For example, the added amount of the culture medium may be 50% - 75% of the total volume of the culture space 10. In one exemplary case, the added volume of the culture medium is 700 mL - 1200 mL, but the present invention is not limited thereto.

[0038] According to experimental measurements, when the average area of the breathable surface S per unit volume of the culture medium is 0.9 cm 2 / mL - 1.9 cm 2 / mL, a balance can be achieved between cell viability and the size of the cell culture bag. For example, the average area of the breathable surface S per unit volume of the culture medium is 1.0 cm 2 / mL, 1.2 cm 2 / mL, 1.4 cm 2 / mL, 1.6 cm 2 / mL or 1.8 cm 2 / mL.

[0039] For example, in one exemplary case, the total area of the breathable surfaces S on both sides of the plastic bag 1 is 1314 cm 2 . When the added amount of the culture medium is 700 mL, the average area of the breathable surface S per unit volume of the culture medium is 1.877 cm 2 / mL. When the added amount of the culture medium is 1200 mL, the average area of the breathable surface S per unit volume of the culture medium is 1.095 cm 2 / mL.

[0040] To improve the breathability of the plastic bag 1, the material of the plastic bag 1 may include at least one selected from the group consisting of ethylene - vinyl acetate copolymer (EVA), linear low density polyethylene (LLDPE), and low density polyethylene (LDPE).

[0041] Specifically, the melting point of the ethylene-vinyl acetate copolymer by differential scanning calorimetry (DSC) is 80°C to 90°C, preferably 82°C to 86°C. The weight-average molecular weight of the ethylene-vinyl acetate copolymer by gel permeation chromatography (GPC) is 31,000 g / mol to 33,000 g / mol, preferably 31,500 g / mol to 32,500 g / mol. The melt index of the ethylene-vinyl acetate copolymer by the ASTM D1238 standard measurement method is 1.6 to 2.0, preferably 1.7 to 1.9. The density of the ethylene-vinyl acetate copolymer by the ASTM D1505 standard measurement method is 0.910 g / cm 3 ~0.950 g / cm 3 and preferably 0.935 g / cm 3 ~0.940 g / cm 3 .

[0042] Specifically, the melting point of the linear low-density polyethylene by differential scanning calorimetry is 116°C to 125°C, preferably 120°C to 124°C. The weight-average molecular weight of the linear low-density polyethylene by gel permeation chromatography is 132,000 g / mol to 134,000 g / mol, preferably 132,500 g / mol to 133,500 g / mol. The melt index of the linear low-density polyethylene by the ASTM D1238 standard measurement method is 1.8 to 2.2, preferably 1.9 to 2.1. The density of the linear low-density polyethylene by the ASTM D1505 standard measurement method is 0.910 g / cm 3 ~0.950 g / cm 3 and preferably 0.915 g / cm 3 ~0.925 g / cm 3 .

[0043] Specifically, the melting point of low-density polyethylene measured by differential scanning calorimetry is 106°C to 115°C, preferably 109°C to 113°C. The weight-average molecular weight of low-density polyethylene by gel permeation chromatography is 129,000 g / mol to 131,000 g / mol, and the weight-average molecular weight of low-density polyethylene is preferably 129,500 g / mol to 130,500 g / mol. The melt index of low-density polyethylene by the ASTM D1238 standard measurement method is 1.8 to 2.2, and the melt index of low-density polyethylene is preferably 1.9 to 2.1. The density of low-density polyethylene by the ASTM D1505 standard measurement method is 0.910 g / cm 3 ~0.950 g / cm 3 and the density of low-density polyethylene is preferably 0.920 g / cm 3 ~0.925 g / cm 3 .

[0044] As shown in FIG. 2, the thickness T of the plastic bag 1 also affects the air permeability. When the thickness T of the plastic bag 1 is 50 μm to 140 μm, the plastic bag 1 has good air permeability. Specific experimental explanations are as follows.

[0045] By selecting materials and adjusting the thickness, the oxygen permeability of the plastic bag 1 at 37°C and 1 atm is more than 200 g / m 2 / day, and the water vapor permeability of the plastic bag 1 at 37°C and 1 atm is less than 10 g / m 2 / day.

[0046] To prove that the cell culture bag according to the present invention has good air permeability, plastic bags with different thicknesses (Experimental Examples 1 to 9) were manufactured using ethylene vinyl acetate copolymer, linear low-density polyethylene, and low-density polyethylene, respectively. In addition, the characteristics of the plastic bags were measured, and the measurement results are as shown in Table 1.

[0047] In Table 1, the water vapor transmission rate of the plastic bag was measured in an environment of 37°C and 1 atm based on the ASTM F1249 standard measurement method. The oxygen transmission rate of the plastic bag was measured in an environment of 37°C and 1 atm based on the ASTM D3985 standard measurement method. Also, the light transmittance and haze value of the plastic bag were measured with a haze meter (ASTM D1003), the surface resistance value of the plastic bag was measured with a surface resistance meter, and the tensile strength of the plastic bag was measured with a universal tensile tester (ASTM D882).

[0048]

Table 1

[0049] According to the results in Table 1, when the thickness of the plastic bag was too thick, the oxygen transmission rate of the plastic bag decreased, and the desired oxygen transmission rate (exceeding 200 g / m 2 / day) could not be achieved. When the thickness of the plastic bag was too thin, the oxygen transmission rate of the plastic bag was sufficient for cell growth, but the water vapor transmission rate of the plastic bag also increased accordingly. When the water vapor transmission rate was too fast or the concentration of the culture solution was too high, it led to cell death. Therefore, in the comprehensive evaluation of the oxygen transmission rate and the water vapor transmission rate, the thickness of the plastic bag is preferably 50 μm to 140 μm.

[0050] According to the results in Table 1, by adopting ethylene vinyl acetate copolymer, linear low density polyethylene or low density polyethylene as the material of the plastic bag, an appropriate oxygen transmission rate and water vapor transmission rate were given to the plastic bag, and an appropriate light transmittance (exceeding 90%) and haze value (less than 11%) were given to the plastic bag.

[0051] In addition, the tensile strength of the plastic bag exceeds 3 kgf, and preferably, the tensile strength of the plastic bag is 3 kgf to 5.5 kgf. In this way, breakage due to the action of external force can be avoided in the processing operation of the plastic bag. Usually, the growth state of cells is inferred by measuring the resistance value of the cell culture bag. Therefore, the surface resistance value of the plastic bag needs to exceed 2×10 12 to avoid the measurement result of the plastic bag.

[0052] The cell culture bag according to another embodiment of the present invention includes a structure similar to the cell culture bag in FIG. 1. The cell culture bag includes a plastic bag 1, a feed connection part 2, a sampling connection part 3, and a discharge connection part 4 installed on the plastic bag 1. The difference is that the plastic bag 1 is made of a two-layer film.

[0053] As shown in FIG. 3, after the two-layer film is processed by a heat-sealing process, the parts of the two-layer film processed by the heat-sealing process are joined to each other by high-temperature melting to form a sheet part 11 in the plastic bag 1. The sheet part 11 defines the range of the culture space 10 so as to surround the air-permeable surface S. In one example, the thickness of the two-layer film is 50 μm to 140 μm.

[0054] Specifically, the two-layer film includes a first layer 101 and a second layer 102, and the first layer 101 is installed on the second layer 102. The second layer 102 is inside the plastic bag 1, that is, the second layer 102 faces the culture space 10. Therefore, in the heat-sealing process, the second layer 102 is the main part that forms the sheet part 11 by fusion.

[0055] To achieve the ideal effect, the thickness T of the two-layer film shall not exceed 150 μm. Therefore, in the heat-sealing process, the two-layer film is prone to film breakage. To solve this problem, in the present invention, films with different melting points are laminated to produce a two-layer film, where the melting point of the first layer 101 is higher than that of the second layer 102. Thus, in the heat-sealing process, the sheet portion 11 can be formed at a relatively low heat-sealing temperature, avoiding film breakage.

[0056] Due to the difference in melting points, the two-layer film has good air permeability and a low processing temperature. In one example, the material of the first layer 101 is linear low-density polyethylene, and the material of the second layer 102 is low-density polyethylene. Since the material types of the first layer 101 and the second layer 102 are similar, the two-layer film can be produced by coextrusion. The main difference between the first layer 101 and the second layer 102 is the difference in melting points. The melting point of the first layer 101 may be 3°C to 20°C higher than that of the second layer 102. Preferably, the melting point of the first layer 101 is 5°C to 15°C higher than that of the second layer 102.

[0057] According to experimental measurements, when the value of the ratio of the thickness of the first layer 101 to the thickness of the second layer 102 is 6 to 12, the two-layer film can achieve both air permeability and good heat-sealing effect. Preferably, the value of the ratio of the thickness of the first layer 101 to the thickness of the second layer 102 is 8 to 10.

[0058] To prove that the cell culture bag according to the present invention has good air permeability, after manufacturing a 100-μm two-layer film by coextrusion of linear low-density polyethylene and low-density polyethylene, plastic bags (Experimental Examples 10 to 12) were manufactured using the two-layer film. The two-layer films in Experimental Examples 10 to 12 have a first layer 101 and a second layer 102 with different thickness ratios.

[0059] In addition, the water vapor transmission rate, oxygen transmission rate, light transmission rate, haze value, surface resistance value, tensile strength, and heat seal bag breakage rate of the plastic bag were measured, and the measurement results are as shown in Table 2. Here, since the measurement methods of the water vapor transmission rate, oxygen transmission rate, light transmission rate, haze value, surface resistance value, and tensile strength are as described above, they will not be repeatedly described here.

[0060] In the measurement of the heat seal bag breakage rate, a cell culture bag was manufactured by heat-sealing the plastic bag at a temperature of 115°C. The cell culture bag was filled with 70% liquid, and a 1 kg heavy object was placed on the cell culture bag to observe whether the liquid oozed out from the cell culture bag. A plurality of plastic bags were each measured for liquid containment and placing a heavy object, and the number of times was counted to express the breakage rate of the cell culture bag as a ratio. For example, when the breakage rate is 15%, it indicates that the breakage of the cell culture bag occurred 3 times in 20 measurements.

[0061]

Table 2

[0062] According to the results in Table 2, when the value of the thickness ratio of the first layer to the second layer is 9:1, the cell culture bag has the lowest heat seal bag breakage rate. Also, the heat seal bag breakage rate of the cell culture bag increased along with the increase in the thickness ratio of the second layer.

[0063] According to the results in Table 1 and Table 2, on the premise that the cell culture bag has the desired oxygen transmission rate and water vapor transmission rate due to the installation of the first layer and the second layer, an excellent heat seal effect can be achieved, and the breakage of the bag is less likely to occur.

[0064] [Advantageous Effects according to the Embodiment] As an advantageous effect of the present invention, the cell culture bag according to the present invention has "selection of the material of the plastic bag" and "the average area of the breathable surface per unit volume of the culture space is 0.6 cm2 / mL to 1.5 cm 2The technical feature of "being [value] / mL" gives the cell culture bag good oxygen permeability and appropriate water vapor permeability.

[0065] Furthermore, in order to improve the production yield of the cell culture bag while maintaining appropriate air permeability, the cell culture bag can be manufactured with a two-layer film. By laminating two layers with different melting points, the processing temperature in the heat-sealing process can be reduced, and it is possible to avoid the two-layer film breaking the film in the heat-sealing process.

[0066] The content disclosed above is only a preferred feasible embodiment of the present invention, and the scope of the claims of the present invention is not limited thereto. Therefore, all equivalent technical changes made using the content of the specification and drawings of the present invention are included in the scope of the claims of the present invention.

Explanation of Reference Numerals

[0067] 1... Plastic bag 2... Salary connection part 3... Sampling connection part 4... Discharge connection part 10... Culture space 11... Sheet part W... Width 12.. Leak prevention part S... Air-permeable surface T... Thickness 101... First layer 102... Second layer

Claims

1. A cell culture bag comprising a plastic bag body including a culture space and a breathable surface, and at least two connection parts installed on the plastic bag body, wherein the culture space and the external environment can be in liquid communication through the connection parts, and the plastic bag body is made of a two-layer film containing polyolefin, the two-layer film includes a first layer and a second layer, and the melting point of the first layer is higher than that of the second layer, the culture space is formed inside the plastic bag, and the second layer faces the culture space, the culture space and the external environment are in gas communication through the breathable surface, The average area of the air-permeable surface per unit volume of the culture space is 0.6 cm ２ / mL to 1.5 cm ２ / mL, and The oxygen permeability of the cell culture bag in an environment of 37 °C and 1 atm is more than 200 g / m ２ / day, and the water vapor permeability is less than 10 g / m ２ / day, and the cell culture bag is characterized by this.

2. The cell culture bag according to claim 1, wherein the thickness of the two-layer film is 50 μm to 140 μm.

3. The cell culture bag according to claim 1, wherein the value of the ratio of the thickness of the first layer to the second layer (first layer / second layer) is 6 to 12.

4. The cell culture bag according to claim 1, wherein the plastic bag includes a sheet part formed by heat-sealing to the second layer, and the sheet part defines the range of the culture space so as to surround the breathable surface.

5. The cell culture bag according to claim 4, wherein the width of the sheet part is 6 mm to 15 mm.

6. The cell culture bag according to claim 1, wherein 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, wherein the weight average molecular weight of the linear low-density polyethylene is 132,000 g / mol to 134,000 g / mol.

8. The cell culture bag according to claim 1, wherein 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, wherein the weight average molecular weight of the low-density polyethylene is 129,000 g / mol to 131,000 g / mol.

10. The culture space contains a culture solution, and the average area of the air-permeable surface per unit volume of the culture solution is 1.0 cm ２ / mL to 1.9 cm ２ / mL. The cell culture bag according to claim 1.

11. The cell culture bag according to claim 10, wherein the total volume of the culture space is 1000 mL to 2000 mL, and the total volume of the culture solution is 700 mL to 1200 mL.

12. A cell culture bag comprising a plastic bag body including a culture space and a breathable surface, and at least two connection parts installed on the plastic bag body, The culture space and the external environment can be in liquid communication through the connection part, and the material of the plastic bag is selected from the group consisting of ethylene vinyl acetate copolymer, linear low density polyethylene, and low density polyethylene. The culture space is formed inside the plastic bag, and the culture space and the external environment are in gas communication through the air-permeable surface. The average area of the air-permeable surface per unit volume of the culture space is 0.6 cm ２ / mL to 1.5 cm ２ / mL, and the oxygen permeability of the cell culture bag at 37 °C and 1 atm is more than 200 g / m ２ / day, and the water vapor permeability is less than 10 g / m ２ / day. A cell culture bag characterized by this.

13. The cell culture bag according to claim 12, wherein the material of the plastic bag is an ethylene vinyl acetate copolymer having a melting point of 80°C to 90°C.

14. The cell culture bag according to claim 13, wherein the weight average molecular weight of the ethylene vinyl acetate copolymer is 31,000 g / mol to 33,000 g / mol.

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