Chicken myoblast culture method based on parallel bioreactor

Abstract

The invention discloses a chicken myoblast culture method based on a parallel bioreactor, and belongs to the technical field of cell biology. The chicken myoblast culture method specifically comprises the following steps: (1) transferring a chicken myoblast suspension into a centrifugal tube, and centrifuging in a centrifugal machine; (2) after centrifuging, sucking and removing supernate in the centrifugal tube, and adding a fresh culture medium to resuspend the cells; (3) subculturing the chicken myoblasts; (4) inoculating into a bioreactor, adding a microcarrier and culture medium mixed solution, and continuously culturing; and (5) after ending, counting and harvesting cells. Wherein the used bioreactor is a quadruple parallel stirring type bioreactor, and the equipment accurately manages culture conditions and greatly reduces time and energy cost, so that the production efficiency and the economic benefit are improved. By combining the microcarrier, the serine and the fish skin collagen, the culture environment of the chicken myoblasts is remarkably optimized, finally, the amplification efficiency and the culture yield of the cells are remarkably improved, and the requirement of large-scale production is met.

Description

Technical Field

[0001] The invention relates to the technical field of cell biology, in particular to a chicken myoblast culture method based on a parallel bioreactor. Background Art

[0002] Cell-cultured meat, also known as lab-grown meat or clean meat, is an emerging technology developed to address environmental and health issues caused by traditional meat production. Traditional animal husbandry not only causes greenhouse gas emissions and consumption of land resources, but is also accompanied by animal welfare and food safety issues. Cell-cultured meat is produced by culturing animal cells in vitro, without involving the slaughter process, aiming to reduce environmental impact and improve food safety. Its production process includes four key steps: cell acquisition, development of serum-free culture medium, large-scale expansion and differentiation of cells, and food safety assessment.

[0003] In the technical solution of using bioreactor for chicken myoblast culture, the prior art has the following major defects:

[0004] (1) Low cell expansion efficiency. In the existing technical solutions, the expansion efficiency of chicken myoblasts is low, resulting in slow cell proliferation, long production cycle and insufficient output. This shortcoming not only increases the time cost of research and production, but also limits the feasibility of large-scale application. The low expansion efficiency is mainly due to the limitations of traditional two-dimensional culture methods. In addition, there is also unevenness in nutrient supply and waste removal in traditional culture methods, resulting in an unsatisfactory cell growth environment, which further inhibits the cell proliferation rate.

[0005] (2) The operation steps are complicated, and the time and economic costs are high. The existing technical solutions require multiple manual operations, which are complicated and prone to operational errors. Frequent operations not only consume a lot of manpower and time, but also significantly increase the overall cost and reduce production efficiency and economic benefits.

[0006] (3) Poor stability and consistency of the culture environment. In the existing technical solutions, the parameters of the culture environment are not precisely controlled, resulting in unstable cell growth effects. The volatility and unpredictability of culture conditions will lead to poor repeatability of experimental results, affect the quality and consistency of cell culture, and limit its potential for application in large-scale production. Summary of the invention

[0007] The object of the present invention is to provide a chicken myoblast culture method based on a parallel bioreactor to solve the problems in the prior art such as low cell proliferation efficiency, complicated operation steps, high time and economic costs, and poor stability and consistency of the culture environment.

[0008] To achieve the above object, the present invention provides the following technical solution: a chicken myoblast culture method based on a parallel bioreactor,

[0009] The chicken myoblast culture method based on parallel bioreactors comprises the following steps:

[0010] S1: Transfer the cell suspension of chicken myoblasts into a centrifuge tube and centrifuge in a centrifuge;

[0011] S2: removing the supernatant in the centrifuge tube, and adding fresh culture medium to resuspend the chicken myoblasts;

[0012] S3: inoculating the resuspended chicken myoblasts into a culture dish for expansion, and digesting and counting the cells after the cells reach a target density;

[0013] S4: digesting the chicken myoblasts described in S3 and inoculating them into a bioreactor, adding a mixture of microcarriers and a culture medium, and adjusting the culture temperature, stirring speed, pH value and dissolved oxygen value of the bioreactor, and culturing for 7 days;

[0014] S5: After the culture is completed, the stirring is stopped, the cells are allowed to settle, the supernatant is removed, the cells on the microcarriers are digested with trypsin, and the cells are counted and harvested;

[0015] In S4, the culture medium mixture includes culture medium, serine and fish skin collagen.

[0016] Preferably, the cell suspension of the chicken myoblasts in step S1 is transferred to a centrifuge tube, and the specification of the centrifuge tube is 15 mL.

[0017] Preferably, the centrifugal conditions of the centrifuge in step S1 are a rotation speed of 1000 rpm and a centrifugal time of 3 min.

[0018] Preferably, the target density of chicken myoblast expansion in step S3 is 1×10 5 Pieces / mL.

[0019] Preferably, the bioreactor in step S4 is a quadruple parallel stirred bioreactor.

[0020] Preferably, in step S4, the concentration of serine is maintained at 20-50 mg / mL throughout the entire culture process.

[0021] Preferably, in step S4, the concentration of the fish skin collagen is maintained at 80-120 mg / mL throughout the entire culture process.

[0022] Preferably, in step S4, the concentration of the microcarriers is maintained at 2 mg / mL throughout the entire culture process.

[0023] Preferably, in step S4, the culture temperature of the bioreactor is 37-41°C; the stirring speed of the bioreactor is 80-120 rpm; the pH value of the bioreactor is 7.2-7.4; and the dissolved oxygen value of the bioreactor is 40%-60%.

[0024] Preferably, in steps S2 and S4, the culture medium is DMEM medium containing 10% FBS.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] ① The present invention adopts parallel bioreactors combined with microcarriers, serine, fish skin collagen and other additives. This technical solution significantly optimizes the culture environment of chicken myoblasts, increases the growth surface area and three-dimensional culture space of the cells, ensures that the cells obtain sufficient nutrition and attachment support, and ultimately significantly improves the cell expansion efficiency and culture yield, meeting the needs of large-scale production.

[0027] ② The present invention reduces the cumbersome manual operation steps in traditional culture methods through a parallel bioreactor system, reduces the possibility of operational errors, and significantly reduces time and energy costs, thereby improving production efficiency and economic benefits.

[0028] ③ Through the precise control of parallel bioreactors, the present invention can monitor and adjust key parameters such as temperature, pH, dissolved oxygen and stirring speed during the culture process in real time, ensuring the stability and consistency of the cell culture environment, thereby improving the repeatability of experimental results and the quality of cell products, and providing reliable technical support for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is a schematic diagram of the process of the chicken myoblast culture method of Example 2 of the present invention;

[0030] Figure 2 The figure is a schematic flow chart of a chicken myoblast culture method according to a comparative example of the present invention;

[0031] Figure 3 Graphs showing cell attachment rates of Examples 1, 2, 3, and 4 of the present invention;

[0032] Figure 4 The cell proliferation curves of Examples 1, 2, 3, and 4 of the present invention are shown;

[0033] Figure 5 Graphs showing cell attachment rates of Examples 4, 5 and 6 of the present invention;

[0034] Figure 6 The cell proliferation curves of Examples 4, 5 and 6 of the present invention are shown;

[0035] Figure 7 This is a comparative example of the present invention, a graph showing the cell attachment rate of Example 4 and a control group without additives (without lysine and leucine);

[0036] Figure 8 The cell proliferation curves of Example 4 and the control group without additives (without lysine and leucine) are comparative examples of the present invention. DETAILED DESCRIPTION

[0037] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. The experimental methods described in the following embodiments are conventional methods unless otherwise specified; the reagents and materials, unless otherwise specified, can be obtained from commercial channels.

[0038] In the following examples 1 to 6, and in the comparative examples, the chicken myoblast recovery method is to take out the chicken myoblast cryopreservation tube from liquid nitrogen, place it in a 37°C water bath to thaw, and wipe the cryopreservation tube mouth with alcohol after taking it out; transfer the cell suspension of the chicken myoblast to a 15mL centrifuge tube and centrifuge it in a centrifuge, the centrifugation condition is that the speed is 1000rpm and the centrifugation time is 3min; after centrifugation, the supernatant in the centrifuge tube is sucked off, and fresh culture medium is added to resuspend the cells. The culture medium used is DMEM culture medium containing 10% FBS.

[0039] Example 1

[0040] Step 1: Inoculate the revived chicken myoblasts into a culture dish and expand them until the cells reach an appropriate density of 1×10 5 After 100 cells / mL, the cells were digested and counted.

[0041] Step 2: Place 1×10 7 The cells were inoculated into parallel bioreactors (the bioreactor was a quadruple parallel stirred bioreactor), and microcarriers and culture medium were added, wherein the culture medium was DMEM culture medium containing 10% FBS. The volume of the culture medium was always 100 mL, and the concentration of the microcarriers was always maintained at 2 mg / mL during the entire culture process. The culture temperature of the bioreactor was precisely controlled to be 37°C, the stirring speed of the bioreactor culture was 120 rpm, the pH value of the bioreactor culture was 7.2, and the dissolved oxygen value of the bioreactor culture was 60%, ensuring the stability and consistency of the culture environment. The cell attachment effect was detected after 24 hours of culture, and the cell proliferation status was detected every other day. The culture process lasted for 7 days, and various parameters were monitored in real time to ensure that the cells grew under optimal conditions.

[0042] Step 3: After the culture is completed, stop stirring, let the culture stand and settle, remove the supernatant and digest the cells on the microcarriers with trypsin. Finally, count and harvest the cells.

[0043] Example 2

[0044] Step 1: Inoculate the revived chicken myoblasts into a culture dish and expand them until the cells reach an appropriate density of 1×10 5 After 100 cells / mL, the cells were digested and counted.

[0045] Step 2: Place 1×10 7 The cells were inoculated into a parallel bioreactor (the bioreactor was a quadruple parallel stirred bioreactor), and a mixture of microcarriers and culture medium was added, and the volume of the culture medium mixture was always 100 mL. The culture medium mixture included serine and culture medium. The culture medium was DMEM culture medium containing 10% FBS. The concentration of microcarriers was maintained at 2 mg / mL throughout the culture process, and the concentration of serine was maintained at 30 mg / mL throughout the culture process to provide a larger surface area, rich nutrients and a suitable extracellular matrix environment. The culture volume of the bioreactor was strictly maintained to ensure the stability of the concentration of the additives. The culture temperature of the bioreactor was precisely controlled to be 37°C, the stirring speed of the bioreactor culture was 120 rpm, the pH value of the bioreactor culture was 7.2, and the dissolved oxygen value of the bioreactor culture was 60%, ensuring the stability and consistency of the culture environment. The cell attachment effect was detected after 24 hours of culture, and the cell proliferation status was detected every other day. The culture process lasted for 7 days, and various parameters were monitored in real time to ensure that the cells grew under optimal conditions.

[0046] Step 3: After the culture is completed, stop stirring, let the culture stand and settle, remove the supernatant and digest the cells on the microcarriers with trypsin. Finally, count and harvest the cells.

[0047] Example 3

[0048] Step 1: Inoculate the revived chicken myoblasts into a culture dish and expand them until the cells reach an appropriate density of 1×10 5 After 100 cells / mL, the cells were digested and counted.

[0049] Step 2: Place 1×10 7Each cell was inoculated into a parallel bioreactor (the bioreactor was a quadruple parallel stirred bioreactor), and a mixture of microcarriers and culture medium was added, and the volume of the culture medium mixture was always 100 mL. The culture medium mixture included fish skin collagen and culture medium. The culture medium was DMEM culture medium containing 10% FBS. The concentration of microcarriers was maintained at 2 mg / mL throughout the culture process. The concentration of fish skin collagen was maintained at 100 mg / mL throughout the culture process to provide a larger surface area, rich nutrients and a suitable extracellular matrix environment. The culture volume of the bioreactor was strictly maintained to ensure the stability of the concentration of the additives. The culture temperature of the bioreactor was precisely controlled to be 37°C, the stirring speed of the bioreactor culture was 120 rpm, the pH value of the bioreactor culture was 7.2, and the dissolved oxygen value of the bioreactor culture was 60%, ensuring the stability and consistency of the culture environment. The cell attachment effect was detected after 24 hours of culture, and the cell proliferation status was detected every other day. The culture process lasted for 7 days, and various parameters were monitored in real time to ensure that the cells grew under optimal conditions.

[0050] Step 3: After the culture is completed, stop stirring, let the culture stand and settle, remove the supernatant and digest the cells on the microcarriers with trypsin. Finally, count and harvest the cells.

[0051] Example 4

[0052] Step 1: Inoculate the revived chicken myoblasts into a culture dish and expand them until the cells reach an appropriate density of 1×10 5 After 100 cells / mL, the cells were digested and counted.

[0053] Step 2: Place 1×10 7Each cell was inoculated into a parallel bioreactor (the bioreactor was a quadruple parallel stirred bioreactor), and a mixture of microcarriers and culture medium was added, and the volume of the culture medium mixture was always 100 mL. The culture medium mixture included serine, fish skin collagen and culture medium. The culture medium was DMEM culture medium containing 10% FBS. The concentration of microcarriers was always maintained at 2 mg / mL throughout the culture process. The concentration of serine was always maintained at 30 mg / mL throughout the culture process. The concentration of fish skin collagen was always maintained at 100 mg / mL throughout the culture process to provide a larger surface area, rich nutrients and a suitable extracellular matrix environment. The culture volume of the bioreactor was strictly maintained to ensure the stability of the concentration of the additives. The culture temperature of the bioreactor was precisely controlled to be 37°C, the stirring speed of the bioreactor culture was 120 rpm, the pH value of the bioreactor culture was 7.2, and the dissolved oxygen value of the bioreactor culture was 60%, ensuring the stability and consistency of the culture environment. The cell attachment effect was detected after 24 hours of culture, and the cell proliferation status was detected every other day. The culture process lasted for 7 days, and various parameters were monitored in real time to ensure that the cells grew under optimal conditions.

[0054] Step 3: After the culture is completed, stop stirring, let the culture stand and settle, remove the supernatant and digest the cells on the microcarriers with trypsin. Finally, count and harvest the cells.

[0055] Example 5

[0056] Step 1: Inoculate the revived chicken myoblasts into a culture dish and expand them until the cells reach an appropriate density of 1×10 5 After 100 cells / mL, the cells were digested and counted.

[0057] Step 2: Place 1×10 7Each cell was inoculated into a parallel bioreactor (the bioreactor was a quadruple parallel stirred bioreactor), and a mixture of microcarriers and culture medium was added, and the volume of the culture medium mixture was always 100 mL. The culture medium mixture included serine, fish skin collagen and culture medium. The culture medium was DMEM culture medium containing 10% FBS. The concentration of microcarriers was always maintained at 2 mg / mL throughout the culture process. The concentration of serine was always maintained at 20 mg / mL throughout the culture process. The concentration of fish skin collagen was always maintained at 120 mg / mL throughout the culture process to provide a larger surface area, rich nutrients and a suitable extracellular matrix environment. The culture volume of the bioreactor was strictly maintained to ensure the stability of the concentration of the additives. The culture temperature of the bioreactor was precisely controlled to be 41°C, the stirring speed of the bioreactor culture was 100 rpm, the pH value of the bioreactor culture was 7.4, and the dissolved oxygen value of the bioreactor culture was 40%, ensuring the stability and consistency of the culture environment. The cell attachment effect was detected after 24 hours of culture, and the cell proliferation status was detected every other day. The culture process lasted for 7 days, and various parameters were monitored in real time to ensure that the cells grew under optimal conditions.

[0058] Step 3: After the culture is completed, stop stirring, let the culture stand and settle, remove the supernatant and digest the cells on the microcarriers with trypsin. Finally, count and harvest the cells.

[0059] Example 6

[0060] Step 1: Inoculate the revived chicken myoblasts into a culture dish and expand them until the cells reach an appropriate density of 1×10 5 After 100 cells / mL, the cells were digested and counted.

[0061] Step 2: Place 1×10 7Each cell was inoculated into a parallel bioreactor (the bioreactor was a quadruple parallel stirred bioreactor), and a mixture of microcarriers and culture medium was added, and the volume of the culture medium mixture was always 100 mL. The culture medium mixture included serine, fish skin collagen and culture medium. The culture medium was DMEM culture medium containing 10% FBS. The concentration of microcarriers was always maintained at 2 mg / mL throughout the culture process. The concentration of serine was always maintained at 50 mg / mL throughout the culture process. The concentration of fish skin collagen was always maintained at 80 mg / mL throughout the culture process to provide a larger surface area, rich nutrients and a suitable extracellular matrix environment. The culture volume of the bioreactor was strictly maintained to ensure the stability of the concentration of the additives. The culture temperature of the bioreactor was precisely controlled to be 38°C, the stirring speed of the bioreactor culture was 80 rpm, the pH value of the bioreactor culture was 7.3, and the dissolved oxygen value of the bioreactor culture was 50%, ensuring the stability and consistency of the culture environment. The cell attachment effect was detected after 24 hours of culture, and the cell proliferation status was detected every other day. The culture process lasted for 7 days, and various parameters were monitored in real time to ensure that the cells grew under optimal conditions.

[0062] Step 3: After the culture is completed, stop stirring, let the culture stand and settle, remove the supernatant and digest the cells on the microcarriers with trypsin. Finally, count and harvest the cells.

[0063] Comparative Example

[0064] Step 1: Inoculate the revived chicken myoblasts into a culture dish and expand them until the cells reach an appropriate density of 1×10 5 After 100 cells / mL, the cells were digested and counted.

[0065] Step 2: Place 1×10 7Each cell was inoculated into a parallel bioreactor (the bioreactor was a quadruple parallel stirred bioreactor), and a microcarrier and a culture medium mixture were added, and the volume of the culture medium mixture was always 100 mL. The culture medium mixture included lysine, leucine and culture medium. The culture medium was a DMEM culture medium containing 10% FBS. The concentration of the microcarrier was maintained at 2 mg / mL throughout the culture process. The concentration of lysine was maintained at 1 mg / mL throughout the culture process and the concentration of leucine was maintained at 0.5 mg / mL throughout the culture process to provide a larger surface area, rich nutrients and a suitable extracellular matrix environment. The culture volume of the bioreactor was strictly maintained to ensure the stability of the concentration of the additives. The culture temperature of the bioreactor was precisely controlled to be 37°C, the stirring speed of the bioreactor culture was 120 rpm, the pH value of the bioreactor culture was 7.2, and the dissolved oxygen value of the bioreactor culture was 60%, ensuring the stability and consistency of the culture environment. The cell attachment effect was detected after 24 hours of culture, and the cell proliferation status was detected every other day. The culture process lasted for 7 days, and various parameters were monitored in real time to ensure that the cells grew under optimal conditions.

[0066] Step 3: After the culture is completed, stop stirring, let the culture stand and settle, remove the supernatant and digest the cells on the microcarriers with trypsin. Finally, count and harvest the cells.

[0067] Figure 1 It is a schematic diagram of the process of the chicken myoblast culture method of Example 2. Figure 2 Schematic diagram of the process of chicken myoblast culture method for comparison.

[0068] Results and Discussion

[0069] (1) Figure 3 : It is the cell attachment rate graph of Examples 1, 2, 3, and 4 of the present invention. The study tested the effects of adding serine, fish skin collagen, and adding these two substances at the same time on the cell attachment rate. Compared with Example 1 without any additives, the cell attachment rate was significantly improved after adding serine and fish skin collagen. This is because serine has a positive promoting effect on the synthesis of extracellular matrix and cell surface proteins, while fish skin collagen provides more adhesion sites for cells. When the two are used at the same time, the cell attachment rate is further improved, showing a synergistic effect.

[0070] (2) Figure 4 It is the cell proliferation curve diagram of Examples 1, 2, 3, and 4 of the present invention. Figure 4The effects of adding serine, fish skin collagen, and both on the cell proliferation efficiency when chicken myoblasts were cultured in parallel bioreactors in Examples 1, 2, 3, and 4 were demonstrated. Compared with Example 1 without any additives, the addition of 30 mg / mL of serine or 100 mg / mL of fish skin collagen significantly improved the cell proliferation efficiency. When serine and fish skin collagen were added, the cell proliferation effect was even more significant. This increase in proliferation efficiency is mainly attributed to the fact that serine and fish skin collagen provide cells with additional surface area, nutrients, and an extracellular matrix environment, which helps cell attachment and proliferation.

[0071] (3) Figure 5 Graph showing cell attachment rates of Examples 4, 5 and 6 of the present invention. Figure 5 The experimental results of culturing chicken myoblasts using parallel bioreactors in Examples 4, 5, and 6 are shown. Each group of experiments was conducted under different culture conditions, with different concentrations of serine and fish skin collagen added, and the parameters of the culture environment were adjusted to investigate the effects of different combinations of conditions on the attachment efficiency of chicken myoblasts. The experimental results show that despite the differences in the concentrations of serine and fish skin collagen, the differences in cell attachment rates between the groups are not significant, indicating that within the concentration range tested, the addition of these two substances has a limited effect on the cell attachment effect.

[0072] (4) Figure 6 Graph showing cell attachment rates of Examples 4, 5 and 6 of the present invention. Figure 6 It is shown that in Examples 4, 5, and 6, when chicken myoblasts are cultured using parallel bioreactors, different concentrations of serine and fish skin collagen are added simultaneously, and the reactors are operated under different culture conditions. The experimental results show that whether it is adjusting the concentration of serine and fish skin collagen or changing the specific culture parameters of the reactor (such as temperature, stirring rate, dissolved oxygen concentration, etc.), the difference in cell proliferation in the three embodiments is not significant. Although the adjustment of different concentrations and conditions has an effect on the cell microenvironment, the final effect on cell proliferation is relatively small. This shows that within a certain range, the changes in the added concentration of serine and fish skin collagen and the culture conditions have no significant difference in the proliferation effect of chicken myoblasts, indicating that these conditions are sufficient to meet the proliferation needs of cells.

[0073] (5) Figure 7 This is a comparative example of the present invention, a graph of cell attachment rate of Example 4 and a control group without additives (without lysine and leucine). Figure 7It was shown that the addition of lysine and leucine in the comparative example significantly improved the cell adhesion effect, and showed a higher adhesion efficiency than the cells without additives in the control group. This shows that lysine and leucine not only provide the necessary amino acid supplements for the cells, but also promote the binding of cells to the microcarrier surface by improving the environment of the extracellular matrix or enhancing the adhesion properties of the cell membrane. However, compared with Example 4, the cell adhesion rate is still low, so it is used as an alternative. In contrast, the effect of lysine and leucine is not as good as the combination of serine and fish skin collagen, which performs better in cell adhesion rate, because the combination of serine and fish skin collagen can better simulate the extracellular matrix and promote cell adhesion, providing a more favorable adhesion environment and signal transduction support. The control group here does not add lysine and leucine, but other conditions are the same.

[0074] (6) Figure 8 It is a comparative example of the present invention, a cell proliferation curve diagram of Example 4 and a control group without additives (without lysine and leucine). Figure 8 It is stated that after adding lysine and leucine, the cell proliferation ability has been significantly improved. Experimental data show that the comparison group with the addition of these two amino acids is significantly better than the control group without additives in terms of proliferation rate and total cell number. Lysine and leucine are not only involved in protein synthesis as essential amino acids, but also play an important role in cell metabolism and signal transduction, thereby promoting cell cycle progression and accelerating cell division and growth. In addition, these two amino acids may enhance the cell's ability to respond to external stimuli and improve the overall culture effect by providing a better nutritional environment. The control group here did not add lysine and leucine, but other conditions were the same.

[0075] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for culturing chicken myoblasts based on parallel bioreactors, characterized in that: The chicken myoblast culture method based on parallel bioreactors comprises the following steps: S1: Transfer the cell suspension of chicken myoblasts into a centrifuge tube and centrifuge in a centrifuge; S2: removing the supernatant in the centrifuge tube, and adding fresh culture medium to resuspend the chicken myoblasts; S3: inoculating the resuspended chicken myoblasts into a culture dish for expansion, and digesting and counting the cells after the cells reach a target density; S4: digesting the chicken myoblasts described in S3 and inoculating them into a bioreactor, adding a mixture of microcarriers and a culture medium, and adjusting the culture temperature, stirring speed, pH value and dissolved oxygen value of the bioreactor, and culturing for 7 days; S5: After the culture is completed, the stirring is stopped, the cells are allowed to settle, the supernatant is removed, the cells on the microcarriers are digested with trypsin, and the cells are counted and harvested; In S4, the culture medium mixture includes culture medium, serine and fish skin collagen.

2. The chicken myoblast culture method based on parallel bioreactor according to claim 1, characterized in that: The cell suspension of the chicken myoblasts in step S1 is transferred to a centrifuge tube, and the specification of the centrifuge tube is 15 mL.

3. The chicken myoblast culture method based on parallel bioreactor according to claim 1, characterized in that: The centrifugal conditions of the centrifuge in step S1 are a rotation speed of 1000 rpm and a centrifugal time of 3 min.

4. The chicken myoblast culture method based on parallel bioreactor according to claim 1, characterized in that: The target density of the chicken myoblast expansion in step S3 is 1×10 5 Pieces / mL.

5. The chicken myoblast culture method based on parallel bioreactor according to claim 1, characterized in that: The bioreactor in step S4 is a quadruple parallel stirred bioreactor.

6. The chicken myoblast culture method based on parallel bioreactor according to claim 1, characterized in that: In step S4, the concentration of serine is maintained at 20-50 mg / mL throughout the culture process.

7. The chicken myoblast culture method based on parallel bioreactor according to claim 1, characterized in that: In step S4, the concentration of the fish skin collagen is always maintained at 80-120 mg / mL during the entire culture process.

8. The chicken myoblast culture method based on parallel bioreactor according to claim 1, characterized in that: In step S4, the concentration of the microcarriers was maintained at 2 mg / mL throughout the culture process.

9. The chicken myoblast culture method based on parallel bioreactor according to claim 1, characterized in that: In step S4, the culture temperature of the bioreactor is 37-41° C.; the stirring speed of the bioreactor is 80-120 rpm; the pH value of the bioreactor is 7.2-7.4; and the dissolved oxygen value of the bioreactor is 40%-60%.

10. The chicken myoblast culture method based on parallel bioreactor according to claim 1, characterized in that: In steps S2 and S4, the culture medium was DMEM medium containing 10% FBS.

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