New-form ginseng stem cell culture system, culture method and device

Abstract

The invention discloses a novel ginseng stem cell culture system which comprises a basic culture medium and a growth regulation composition, and the basic culture medium comprises ammonium sulfate, monopotassium phosphate, magnesium sulfate heptahydrate and calcium chloride; the growth regulating composition comprises 2, 4-dichlorphenoxyacetic acid, zeatin, salicylic acid and natural polysaccharide. The basic culture medium also comprises casein hydrolysate, coconut water extract and cane sugar. The composition also comprises an anti-oxidation composition. The invention further discloses a disposable flexible bioreactor for ginseng stem cell culture, the bioreactor comprises a tank body made of a flexible material, an air distribution device, a sterile sampling device and an online parameter monitoring device, and the top of the tank body structure is a cylindrical body and is provided with an inoculation opening, a culture medium liquid supplementing opening, an air outlet and a sampling opening; the bottom is an inverted cone provided with a parameter monitoring device interface and an air inlet, and the air inlet is connected with a ventilation device and an air flow adjusting device. The invention also discloses application of the culture system and the bioreactor.

Description

Technical Field

[0001] This invention relates to the fields of cell biology and molecular biology, and in particular to a novel ginseng stem cell culture system, culture method and apparatus. Background Technology

[0002] The concept of stem cells was introduced in animal embryology research in the 1960s, defined as "cells with self-renewal and differentiation potential." In plant anatomy, the corresponding stem cells are various meristematic tissues, such as shoot tips, root tips, lateral meristems, and some medullary ray cells. Compared to traditional callus tissue, these meristematic cells have the following three characteristics: First, multiple small vacuoles, high mitochondrial activity, and low aggregation; second, no lignin deposition in the cell wall, and strong shear stress tolerance; third, high activity after cryopreservation, and stable long-term proliferation in suspension culture as single cells. These characteristics lay the biological foundation for establishing uniform, controllable, and scalable plant cell factories.

[0003] With the rapid development of my country's pharmaceutical and functional food industries, the demand for active ingredients in plants is increasing daily. Ginseng, known as the "King of Herbs," is rich in active ingredients—ginsenosides (especially rare saponins Rg3, Rh2, and Rk2)—which possess anti-aging, immune-regulating, and cardiovascular-protective functions. However, traditional ginseng cultivation methods have the following drawbacks: First, the growth cycle is long, requiring ginseng roots to accumulate sufficient ginsenosides only after six years or more; second, the distribution of effective components in the resulting ginseng roots is uneven, greatly affected by soil, climate, and diseases. Therefore, developing a new form of ginseng stem cell culture system to transform this "millennial medicinal plant" into a "modern cell factory" breaks through resource limitations, quality fluctuations, and environmental bottlenecks, providing a sustainable, traceable, and scalable new method for the application of ginseng in high-end functional foods and precision medicine is highly promising. Summary of the Invention

[0004] The purpose of this invention is to provide a novel ginseng stem cell culture system, culture method, and apparatus.

[0005] To achieve the above objectives, the solution of the present invention is:

[0006] A novel ginseng stem cell culture system comprising a basal culture medium and a growth regulating composition, wherein the basal culture medium comprises the following components: ammonium sulfate, potassium dihydrogen phosphate, magnesium sulfate heptahydrate, and calcium chloride; and the growth regulating composition comprises: 2,4-dichlorophenoxyacetic acid, zeatin, salicylic acid, and natural polysaccharides.

[0007] Preferably, the basal culture medium further comprises the following components: hydrolyzed casein, coconut water extract, and sucrose.

[0008] Preferably, it also includes an antioxidant composition.

[0009] A disposable flexible bioreactor for ginseng stem cell culture, characterized in that it comprises:

[0010] The container is made of flexible material, and includes an air distribution device, a sterile sampling device, and an online parameter monitoring device. The top of the container is cylindrical and has an inoculation port, a culture medium replenishment port, an air outlet, and a sampling port. The bottom of the container is an inverted cone and has an interface for the parameter monitoring device and an air inlet. The air inlet is connected to a ventilation device and an air flow regulating device.

[0011] Preferably, the flexible material is composed of one or more of polypropylene, polyethylene, polyvinylidene fluoride, and nylon.

[0012] Preferably, the air outlet is located on the top cover of the tank and is connected to an air respirator and an air regulating valve via a pipeline; the air inlet is located at the bottom of the tank and is connected to an air filter and an air flow regulating valve via a pipeline; the sampling port is located at the bottom of the tank (40-60cm) and is connected to a sterile sampling device via a pipeline.

[0013] Preferably, the bottom of the tank is an inverted cone with an included angle of 30-60 degrees between the two inclined surfaces.

[0014] Preferably, the parameter monitoring device interface is located 25-30cm from the bottom of the inverted cone, and has 3-5 reserved openings.

[0015] Preferably, the air distribution device includes a gas-liquid mixing chamber, the top of which is fitted with a porous panel, the panel having circular holes with a diameter of 0.5-2 mm.

[0016] The aforementioned culture system and the previously described disposable flexible bioreactor are used in the culture of ginseng stem cells, including the following steps:

[0017] (1) Prepare a culture medium specifically for ginseng stem cells;

[0018] (2) Place the ginseng tissue block into a disposable flexible bioreactor for ginseng stem cell culture, and then add ginseng stem cell culture medium for culture. During the culture process, it is necessary to adjust the O2-CO2 supply balance in the reactor, adjust and optimize the culture conditions according to the data transmitted in real time by the reactor parameter monitoring device, and supplement the nutrients lacking in the culture process in a timely manner to increase the saponin content.

[0019] (3) After the culture is completed, the cells in the reactor can be centrifuged and freeze-dried to obtain high-content ginseng stem cells.

[0020] The specific principles of the novel ginseng stem cell culture system and culture method provided by this invention are as follows:

[0021] The novel ginseng stem cell culture system provided by this invention consists of a basal culture medium, a growth regulator composition, and an antioxidant composition. The basal culture medium comprises inorganic and organic nutrients. The organic nutrients utilize a mixed carbon source composed of sucrose and natural polysaccharides, which serves both as a carbon source and an osmotic regulator, promoting rapid stem cell proliferation. Furthermore, the antioxidant composition used in this invention employs a formula combining tea polyphenols and luteolin. This not only enhances the antioxidant properties of tea polyphenols and luteolin but also allows tea polyphenols to promote cell proliferation and maintain stemness, while luteolin promotes further stem cell differentiation. The combination of these two components enables the efficient and targeted accumulation of ginsenosides. This invention also designs a disposable bioreactor for ginseng stem cell culture, made of flexible polymer materials such as polypropylene, polyethylene, polyvinylidene fluoride, or nylon. This effectively solves the problem of cell deposition at the bottom caused by traditional rigid material reactors and also effectively reduces shear force damage to cells.

[0022] The advantages of the novel ginseng stem cell culture system, method, and apparatus provided by this invention are as follows:

[0023] (1) The novel ginseng stem cell culture system provided by the present invention can effectively promote cell proliferation and shorten the culture cycle.

[0024] (2) The novel ginseng stem cell culture system provided by the present invention optimizes the culture conditions so that ginseng stem cells can maintain a strong ability to divide and proliferate, and can form cell types with specific functions.

[0025] (3) The ginseng stem cells obtained by the new ginseng stem cell culture method provided by the present invention have significantly increased content of active ingredients and greatly enhanced medicinal value.

[0026] (4) The disposable flexible bioreactor for ginseng stem cell culture designed in this invention has the advantages of simple and convenient operation, which not only reduces the technical requirements for operators, but also effectively reduces the possibility of misoperation. In addition, the designed bioreactor is made of flexible material, which can not only better adapt to various complex operating environments and is not easily damaged, but also provides flexible operation in terms of reactor sterilization, transportation and cell inoculation culture, and can quickly provide production capacity to deal with emergencies.

[0027] (5) The disposable flexible bioreactor for ginseng stem cell culture designed in this invention has a large ratio of actual material volume to effective internal volume, which can accommodate more biological reactants, thereby improving the efficiency of the biological reaction, reducing reaction time, lowering production costs, and reducing wear on equipment, enabling large-scale production. In addition, the designed reactor can freely replace air distribution devices with different pore sizes and distributions, making it suitable for the suspension culture needs of cell clusters of different sizes and shapes. Attached Figure Description

[0028] Figure 1 This is a schematic diagram of the main structure of the disposable flexible bioreactor for ginseng stem cell culture provided by the present invention;

[0029] Explanation of reference numerals in the attached diagram: 1. Tank body; 2. Inoculation port; 3. Culture medium replenishment port; 4. Air inlet; 5. Air outlet; 6. Sampling port; 7. Parameter monitoring device interface one; 8. Parameter monitoring device interface two; 9. Parameter monitoring device interface three; 10. Air distribution device; 41. Air filter; 42. Air flow regulating valve; 51. Air respirator; 52. Air regulating valve.

[0030] Figure 2 This is a schematic diagram of the air distribution device structure of the disposable flexible bioreactor for ginseng stem cell culture provided by the present invention.

[0031] Explanation of reference numerals in the attached drawings: 101, pagoda head; 102, porous panel; 103, gas-liquid mixing chamber.

[0032] Figure 3 This is a schematic diagram of the aseptic sampling device of the disposable flexible bioreactor for ginseng stem cell culture provided by the present invention.

[0033] Explanation of reference numerals in the attached drawings: 62, aseptic sampling device; 63, connector; 64, aseptic sampling bag.

[0034] Figure 4 This is a schematic diagram of an online parameter monitoring device for a disposable flexible bioreactor for ginseng stem cell culture provided by the present invention;

[0035] Explanation of reference numerals in the attached drawings: 71, online parameter monitoring device; 72, seal; 73, monitoring electrode. Detailed Implementation

[0036] The present invention will now be described in further detail with reference to the accompanying drawings and embodiments. It should also be understood that the following embodiments are only for further illustration of the present invention and should not be construed as limiting the scope of protection of the present invention. Specific mass, reaction time, temperature, process parameters, etc., in the examples are merely examples within a suitable range. Any non-essential improvements and adjustments made by those skilled in the art based on the above description of the present invention are within the scope of protection of the present invention.

[0037] Before use, all bioreactors, inoculation devices, and corresponding auxiliary instruments used in the examples must be disinfected and sterilized. The specific conditions are as follows: auxiliary instruments are sprayed with 75% alcohol and then placed on a clean bench for 20-30 minutes of UV irradiation; inoculation devices are sterilized by moist heat sterilization in a high-pressure steam sterilizer at 121°C and 0.1MPa for 20 minutes; and disposable bioreactors are sterilized by irradiation with a cobalt-60 dose of 10-12 kgy.

[0038] The culture media used in the examples need to be sterilized before use. The specific conditions are as follows: heat-insensitive components can be sterilized by high-temperature and high-pressure steam sterilization at 121°C and 0.1MPa for 20 minutes; heat-sensitive components (zeatin, ascorbic acid, glutathione, luteolin, hydrolyzed casein, tea polyphenols, and natural polysaccharide compounds) should be sterilized by high-pressure steam sterilization first, then cooled to 55–60°C and aseptically added to the culture medium through a 0.22μm filter membrane in a laminar flow hood; after filtration and addition, immediately dispense or pour into plates, place in a 37°C microbial culture room for 24 hours to observe for sterility before use.

[0039] The ginseng explants used in the examples need to be sterilized before use. Taking ginseng root as an example, the specific steps are as follows:

[0040] 1. Select healthy, rot-free, and pest-free ginseng roots as explants;

[0041] 2. Use a soft brush to gently brush the surface of the explant and remove the soil and attachments for 20 minutes under running water; then use scissors that have been pre-sterilized in 75% alcohol to cut the cleaned explant into ginseng root segments of 2–4 cm.

[0042] 3. On a clean bench, immerse the ginseng root segments in 75% ethanol for 30 seconds to quickly kill surface bacteria and fungi; then immediately rinse with sterile water 2-3 times to remove residual ethanol and obtain ethanol-sterilized ginseng root segments.

[0043] 4. Immerse the ginseng root segments disinfected with ethanol in a 2% sodium hypochlorite solution for 20 minutes. Shake the container continuously during disinfection to ensure that the sodium hypochlorite solution is in full contact with the ginseng root segments. After disinfection, rinse with sterile water 4–6 times until no disinfectant residue remains.

[0044] 5. On sterile filter paper, cut off the browned or damaged edges caused by disinfection, and then cut the ginseng root segment into irregular thin slices or small segments with a thickness of 0.2–0.5 mm and a length and width of 5–7 mm. This will give you ginseng tissue blocks for subsequent culture.

[0045] Example 1:

[0046] like Figure 1 The diagram shows the main structure of the disposable flexible bioreactor for ginseng stem cell culture provided by the present invention; it includes: a tank made of flexible material, an air distribution device, a sterile sampling device, and an online parameter monitoring device. The top of the tank is cylindrical and has an inoculation port, a culture medium replenishment port, an air outlet, and a sampling port; the bottom of the tank is an inverted cone and has a parameter monitoring device interface and an air inlet. The air inlet is connected to a ventilation device and an air flow regulating device; the bottom of the tank is also provided with parameter monitoring device interface one, parameter monitoring device interface two, and parameter monitoring device interface three.

[0047] The flexible bioreactor is made of one or more of the following materials: polypropylene, polyethylene, polyvinylidene fluoride, and nylon.

[0048] The air outlet is located on the top cover of the tank and is connected to an air respirator via a pipeline. An air regulating valve is connected to the end of the pipeline.

[0049] The air inlet is located in the center of the tank, 15-20cm from the bottom, and the end of the pipeline is connected to an air filter and an air flow regulating valve.

[0050] The sampling port is located 40-60cm from the bottom of the tank and 15-20cm from the right side. The end of the pipeline has a connector for connecting a sterile sampling device.

[0051] The bottom of the tank is an inverted cone, with the angle between the two inclined surfaces being 30-60 degrees.

[0052] The interface of the parameter monitoring device is located 25-30cm from the bottom of the inverted cone, with 3-5 reserved ports for connecting pH meters, dissolved oxygen electrodes, glucose monitoring electrodes, cell concentration monitoring electrodes, temperature monitoring electrodes, and carbon dioxide monitoring devices.

[0053] The air distribution device includes a gas-liquid mixing chamber with a perforated panel fitted on the top of the chamber. The panel has round holes with a diameter of 0.5-2mm and is connected to a ventilation device through the air inlet of the tank.

[0054] like Figure 2The diagram shows a schematic of another optional air distribution device in the disposable flexible bioreactor for ginseng stem cell culture provided by the present invention. It includes a gas-liquid mixing chamber with a porous panel at its top. The porous panel has multiple uniformly distributed pores, symmetrically arranged in concentric circles, with a diameter of 0.5-2 mm. Furthermore, the air distribution device is divided into three chambers for gas-liquid exchange by multiple annular bubble baffles, thereby increasing the oxygen content in the culture liquid. The air distribution device can be connected to an air supply device via a pagoda-shaped head and the reactor air inlet. This is a further improvement on the air distribution device based on the aforementioned reactor, allowing for adaptation to different cell cultures by replacing the air distribution device.

[0055] like Figure 3 As shown: The disposable flexible bioreactor for ginseng stem cell culture provided by the present invention also includes a sterile sampling device; specifically, it includes: a sterile sampling device, a connector, and a sterile sampling bag; wherein, the sterile sampling device adopts a multi-bag parallel connection and is connected to the tank through a straight connector; after sampling is completed, the sampling pipeline is sterilized by liquid sealing, which can effectively avoid contamination after sampling.

[0056] like Figure 4 As shown: The disposable flexible bioreactor for ginseng stem cell culture provided by the present invention also includes an online parameter monitoring device; the online parameter monitoring device includes a PE insertion seal for connecting to the monitoring device interface to achieve sterile sealing, and also includes an Arc wireless transmission electrode equipped with a transmission chip, which can transmit data wirelessly in real time via Bluetooth.

[0057] Example 2:

[0058] Taking ginseng cell suspension culture as an example, the specific steps are as follows:

[0059] (1) Prepare a culture medium for ginseng stem cells, comprising the following components by mass concentration: ammonium sulfate 681 mg / L, salicylic acid 0.1 mg / L, fucoidan 30 mg / L; hydrolyzed casein 1000 mg / L, coconut water extract 75 ml / L, sucrose 40 g / L; ascorbic acid 1.0 mg / L, glutathione 0.5 mg / L, tea polyphenols 0.5 mg / L, ethylenediamine-N,N′-disuccinic acid 3 mg / L;

[0060] (2) Under the protection of an alcohol lamp flame, 50g of ginseng tissue blocks were placed into a disposable flexible bioreactor through the inoculation port using an inoculation tool. Then, ginseng stem cell-specific culture medium was added through a tubing extending from the culture medium replenishment port for culture. The specific culture conditions were: temperature 24±1℃, light intensity 800-1200 lux, with intermittent light control, specifically, during the 16-hour light period each day, a 5-minute strong light pulse stimulation of 2000-3000 lux was applied every 2 hours. During the culture process, the O2-CO2 supply balance in the reactor could be adjusted by using an air respirator and air regulating valve connected to the air outlet. Oxygen could be supplied to the reactor through an air filter connected to the air inlet, and the air flow regulating valve could be adjusted to control the ventilation ratio at 0.12-0.24 vvm. During the culture period, the culture conditions could be adjusted and optimized based on the real-time data transmitted by the online monitoring device, and nutrients lacking during the culture process could be added as needed. The entire cell suspension culture cycle was 23 days.

[0061] (3) After the culture is completed, the cells in the reactor are separated by gradient centrifugation. The specific conditions are as follows: first, centrifuge at 100×g for 5 minutes to remove debris, then centrifuge at 800×g for 15 minutes to collect the target cells, and finally centrifuge at 1500×g for 3 minutes. The ginseng stem cells obtained by centrifugation are placed in a freeze dryer for freeze drying to obtain high-purity ginseng stem cell products.

[0062] The detection steps for ginsenoside content in the obtained ginseng stem cells are as follows: 5g of the prepared freeze-dried high-purity ginseng stem cell product was accurately weighed and soaked in 50mL of 85% methanol. The mixture was sonicated at 25℃ for 25min, and this sonication was repeated at least 5 times until no ginsenosides were observed in the thin-layer chromatography. The extracts were combined and evaporated to dryness. The residue was dissolved in anhydrous methanol to prepare a methanol solution containing ginsenosides. The ginsenoside content was analyzed using an Agilent 1200 high-performance liquid chromatograph. The results showed that the total ginsenoside content in the obtained ginseng stem cell product was 1.91%.

[0063] Example 3:

[0064] The method for culturing ginseng stem cells is the same as in Example 2, except that the brown algae oligosaccharide in step (1) is replaced with kelp polysaccharide; the rest of the steps are the same as in Example 1.

[0065] The detection steps for the ginsenoside content in the obtained product were as described in Example 2. The results showed that the total ginsenoside content in the obtained ginseng stem cell product was 2.16%.

[0066] Example 4:

[0067] The method for culturing ginseng stem cells is the same as in Example 2, except that the brown algae oligosaccharide in step (1) is replaced with fucoidan; the rest of the steps are the same as in Example 1.

[0068] The detection steps for the ginsenoside content in the obtained product were as described in Example 2. The results showed that the total ginsenoside content in the obtained ginseng stem cell product was 2.34%.

[0069] Example 5:

[0070] The method for culturing ginseng stem cells is the same as in Example 2, except that the brown algae oligosaccharide in step (1) is replaced with Ganoderma lucidum polysaccharide; the rest of the steps are the same as in Example 1.

[0071] The detection steps for the ginsenoside content in the obtained product were as described in Example 2. The results showed that the total ginsenoside content in the obtained ginseng stem cell product was 5.27%.

[0072] Example 6:

[0073] The method for culturing ginseng stem cells is the same as in Example 2, except that the brown algae oligosaccharide in step (1) is replaced with a mixture of Ganoderma lucidum polysaccharide and fucoidan, wherein the mass concentration of Ganoderma lucidum polysaccharide is 10 mg / L and the mass concentration of fucoidan is 20 mg / L; the remaining steps are the same as in Example 1.

[0074] The detection steps for the ginsenoside content in the obtained product were as described in Example 2. The results showed that the total ginsenoside content in the obtained ginseng stem cell product was 6.87%.

[0075] Example 7:

[0076] The ginseng stem cell culture method is the same as in Example 2, except that the tea polyphenol sugar in step (1) is replaced with luteolin; the rest of the steps are the same as in Example 1.

[0077] The detection steps for the ginsenoside content in the obtained product were as described in Example 2. The results showed that the total ginsenoside content in the obtained ginseng stem cell product was 6.52%.

[0078] Example 8:

[0079] The ginseng stem cell culture method is the same as in Example 2, except that the tea polyphenols in step (1) are replaced with a mixture of tea polyphenols and luteolin, wherein the mass concentration of tea polyphenols is 0.4 mg / L and the mass concentration of fucoidan is 20 mg / L. The detection steps for the ginsenoside content in the resulting product are the same as in Example 2. The result shows that the total ginsenoside content in the ginseng stem cell product is 8.14%.

[0080] Example 9:

[0081] The method for culturing ginseng stem cells is the same as in Example 2, except that the brown algae oligosaccharide in step (1) is replaced with a mixture of Ganoderma lucidum polysaccharide and fucoidan, wherein the mass concentration of Ganoderma lucidum polysaccharide is 10 mg / L and the mass concentration of fucoidan is 20 mg / L; the tea polyphenols are replaced with a mixture of tea polyphenols and luteolin, wherein the mass concentration of tea polyphenols is 0.4 mg / L and the mass concentration of luteolin is 0.1 mg / L; the remaining steps are the same as in Example 1.

[0082] The detection steps for the ginsenoside content in the obtained product were as described in Example 2. The results showed that the total ginsenoside content in the obtained ginseng stem cell product was 8.92%.

[0083] Example 10:

[0084] The method for culturing ginseng stem cells is the same as in Example 2, except that the light conditions in step (2) are changed to ordinary light for 16 hours, and intermittent light control is not used. The entire cell suspension culture cycle is changed to 35 days.

[0085] The detection steps for the ginsenoside content in the obtained product were as described in Example 2. The results showed that the total ginsenoside content in the obtained ginseng stem cell product was 1.92%.

Claims

1. A culture system of a new form of ginseng stem cell, characterized in that, The culture system comprises a basal culture medium and a growth regulator composition. The basal culture medium includes the following components: ammonium sulfate, potassium dihydrogen phosphate, magnesium sulfate heptahydrate, and calcium chloride. The growth regulator composition includes: 2,4-dichlorophenoxyacetic acid, zeatin, salicylic acid, and natural polysaccharides.

2. The novel ginseng stem cell culture system according to claim 1, characterized in that, The basal culture medium also contains the following components: hydrolyzed casein, coconut water extract, and sucrose.

3. The novel ginseng stem cell culture system according to claim 1, characterized in that, It also includes antioxidant compositions.

4. A disposable flexible bioreactor for ginseng stem cell culture, characterized in that, include: The container is made of flexible material, and includes an air distribution device, a sterile sampling device, and an online parameter monitoring device. The top of the container is cylindrical and has an inoculation port, a culture medium replenishment port, an air outlet, and a sampling port. The bottom of the container is an inverted cone and has an interface for the parameter monitoring device and an air inlet. The air inlet is connected to a ventilation device and an air flow regulating device.

5. The disposable flexible bioreactor for ginseng stem cell culture according to claim 4, characterized in that, The flexible material is composed of one or more of polypropylene, polyethylene, polyvinylidene fluoride, and nylon.

6. The disposable flexible bioreactor for ginseng stem cell culture according to claim 4, characterized in that, The air outlet is located on the top cover of the tank and is connected to an air respirator and an air regulating valve via a pipeline; the air inlet is located at the bottom of the tank and is connected to an air filter and an air flow regulating valve via a pipeline; the sampling port is located on the tank near the bottom and is connected to a sterile sampling device via a pipeline.

7. The disposable flexible bioreactor for ginseng stem cell culture according to claim 4, characterized in that, The bottom of the tank is an inverted cone, with the angle between the two inclined surfaces being 30-60 degrees.

8. The disposable flexible bioreactor for ginseng stem cell culture according to claim 4, characterized in that, The interface of the parameter monitoring device is located 25-30cm from the bottom of the inverted cone, and has 3-5 reserved openings.

9. The disposable flexible bioreactor for ginseng stem cell culture according to claim 4, characterized in that, The air distribution device includes a gas-liquid mixing chamber, and a perforated panel is fitted on the top of the chamber. The panel has circular holes with a diameter of 0.5-2 mm.

10. The application of the culture system according to any one of claims 1-3 and the disposable flexible bioreactor according to any one of claims 4-9 in ginseng stem cell culture, characterized in that, Specifically, the following steps are included: (1) Prepare a culture medium specifically for ginseng stem cells; (2) Place ginseng stem cells into a disposable flexible bioreactor for ginseng stem cell culture, and then add ginseng stem cell culture medium for culture; during the culture process, it is necessary to adjust the O2-CO2 supply balance in the reactor, adjust and optimize the culture conditions according to the data transmitted in real time by the reactor parameter monitoring device, and supplement the nutrients lacking in the culture process in a timely manner. (3) After the culture is completed, the cells in the reactor are centrifuged and freeze-dried to obtain high-purity ginseng stem cells.

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