Medium additive for MSC (mesenchymal stem cells) and application of medium additive

Abstract

The invention relates to an MSC culture medium additive and application thereof, and relates to the technical field of cell culture. Comprising the following components: 5-13 [mu] g / L of a composite growth factor, 1-3 g / L of a plant component microemulsion, 10-20 mg / L of amino acid, 0.3-1.5 g / L of plant polysaccharide, 0.1-0.2 g / L of hyaluronic acid, 100-150 mg / L of betaine, 40-80 mg / L of vitamin C, 0.6-1.2 mg / L of trace elements, 10-30 mg / L of a natural plant extract, 10-30 mg / L of hydroxypropyl beta-cyclodextrin and 2-4 mmol / L of acetylglutamine. The culture medium additive can effectively promote MSC proliferation and maintain cell viability and differentiation potential.

Description

Technical Field

[0001] The present invention relates to the technical field of cell culture, and particularly to a culture medium additive for MSCs and its application. Background Art

[0002] Mesenchymal stem cells (MSCs) have shown broad application prospects in the fields of regenerative medicine, cell therapy, and cosmetology due to their characteristics such as multi-directional differentiation potential, immunomodulatory function, and easy acquisition. However, during the in vitro expansion culture process of MSCs, problems such as slow proliferation rate, difficulty in maintaining cell activity, and easy loss of differentiation potential are faced, which greatly limits the large-scale culture and clinical application of MSCs.

[0003] The MSC culture medium is the basic guarantee for MSC culture, which can provide a suitable survival environment for MSC cells. Currently, the culture media used for in vitro expansion culture of MSCs usually rely on animal-derived sera (such as FBS) or synthetic growth factors (such as bFGF). Animal-derived sera may introduce pathogens or trigger immune responses, resulting in growth arrest or rapid senescence of MSCs. Traditional growth factor combinations are prone to protein denaturation, leading to fluctuations in cell proliferation efficiency; single-factor regulation is difficult to synchronously maintain the stemness and differentiation potential of stem cells; signal pathway crosstalk caused by non-specific binding of growth factors often leads to an increase in cell heterogeneity. There are also those that choose not to add animal-derived sera and synthetic growth factors during the culture process of existing MSCs, but the culture effect is not ideal, and there are technical defects such as slow cell proliferation rate, low expression, and limited number of proliferated cells obtained.

[0004] To solve the above problems, the common current practice is to incorporate a culture medium additive for MSCs into the MSC culture medium. Although the commercially available culture medium additives for MSCs can promote the growth of MSCs to a certain extent, there are still problems such as unsatisfactory effects, complex components, and high costs. For example, Chinese invention patent CN112941022B discloses a special culture medium for culturing bone marrow mesenchymal stem cells and its application. The culture medium also includes matrix metalloproteinase 2 (MMP2). Experiments have confirmed that the phenotype of BMSCs treated with the MMP2-containing culture medium remains unchanged, but the growth rate significantly increases. Therefore, this MMP2-containing culture medium can be used for the culture of bone marrow-derived MSCs, enabling them to play a more effective role in clinical treatment and cosmetology, etc. However, its component synergy is insufficient, the differentiation potential is limited, and the serum-free culture compatibility, safety, and stability still need to be further improved.

[0005] It can be seen that developing a culture medium additive for MSCs that can effectively promote MSC proliferation, maintain cell activity and differentiation potential has important practical significance. Summary of the Invention

[0006] The object of the present invention is to provide a culture medium additive for MSCs that can effectively promote the proliferation of MSCs, maintain cell viability and differentiation potential, in order to overcome the deficiencies of the prior art.

[0007] To achieve the above object, the technical solution adopted by the present invention is a culture medium additive for MSCs, which comprises the following components: 5-13 μg / L of composite growth factors, 1-3 g / L of plant component microemulsion, 10-20 mg / L of amino acids, 0.3-1.5 g / L of plant polysaccharides, 0.1-0.2 g / L of hyaluronic acid, 100-150 mg / L of betaine, 40-80 mg / L of vitamin C, 0.6-1.2 mg / L of trace elements, 10-30 mg / L of natural plant extracts, 10-30 mg / L of hydroxypropyl-β-cyclodextrin, and 2-4 mmol / L of acetylglutamine.

[0008] Preferably, the composite growth factors are composed of BMP-2, TGF-β1, EGF, and IGF-1 mixed in a mass ratio of (0.5-1.2):(0.3-0.8):(2-4):(3-8).

[0009] Preferably, the preparation method of the plant component microemulsion comprises the following steps: dispersing curcumin, silybin, and epigallocatechin gallate in an organic solvent, adding an emulsifier, and mixing evenly to obtain a mixed solution; mixing the mixed solution with deionized water, and processing it 5-8 times under a pressure of 100-150 MPa by a high-pressure homogenizer to obtain the plant component microemulsion.

[0010] Preferably, the mass ratio of curcumin, silybin, epigallocatechin gallate, organic solvent, and emulsifier is (6-8):(4-6):(3-5):(40-60):(0.65-1.5).

[0011] Preferably, the organic solvent is at least one of absolute ethanol and propylene glycol.

[0012] Preferably, the emulsifier is at least one of poloxamer 188 and Tween 80.

[0013] Preferably, the volume ratio of the mixed solution to deionized water is 1:3.

[0014] Preferably, the amino acids are at least one of proline, glycine, glutamic acid, and arginine.

[0015] Preferably, the plant polysaccharides are at least one of seaweed polysaccharides and dextrans.

[0016] Preferably, the hyaluronic acid has a weight-average molecular weight of 100-500 kDa.

[0017] Preferably, the trace elements are a mixture of Se and Zn in a mass ratio of (0.1 - 0.3):(0.5 - 1).

[0018] Preferably, the natural plant extract is a mixture of green tea polyphenols and rosmarinic acid in a mass ratio of 3:1.

[0019] Another object of the present invention is to provide a method for preparing the medium additive for MSC, comprising the following steps: mixing each component, adding it into deionized water, stirring evenly under the conditions of 4 - 10 °C and in the dark, then filling it into a freeze-drying bottle, pre-freezing at -60 °C for 2 hours, and then drying in a freeze dryer for 25 - 30 h to obtain the medium additive for MSC.

[0020] Preferably, the vacuum degree of the drying is 5 - 10 Pa and the temperature is -40 °C.

[0021] Another object of the present invention is to provide an application of the medium additive for MSC in the culture of MSC cells. The medium additive for MSC is added to a basic cell culture medium for MSC cell culture; the addition amount of the medium additive for MSC is 0.1 - 0.5 g / L.

[0022] Preferably, the basic cell culture medium is DMEM / F12 complete medium.

[0023] Due to the application of the above technical solutions, the present invention has the following beneficial effects: (1) The medium additive for MSC disclosed by the present invention has a simple and easy preparation method, convenient operation control, high preparation efficiency and finished product qualification rate, low dependence on equipment, and is easy to be mass-produced industrially; it is applicable to the in vitro expansion culture of MSC from various sources and has high popularization and application value.

[0024] (2)The culture medium additive for MSCs disclosed by the present invention comprises the following components: 5-13 μg / L of composite growth factors, 1-3 g / L of plant component microemulsion, 10-20 mg / L of amino acids, 0.3-1.5 g / L of plant polysaccharides, 0.1-0.2 g / L of hyaluronic acid, 100-150 mg / L of betaine, 40-80 mg / L of vitamin C, 0.6-1.2 mg / L of trace elements, 10-30 mg / L of natural plant extracts, 10-30 mg / L of hydroxypropyl-β-cyclodextrin, and 2-4 mmol / L of acetylglutamine. Through the mutual cooperation and joint action of each component, the prepared product can effectively promote the proliferation of MSCs, maintain cell activity and differentiation potential; the plant component microemulsion contains curcumin, silybin, and epigallocatechin gallate. The antioxidant signaling pathway activated by curcumin cooperates with the autophagy process triggered by silybin, reducing intracellular oxidative stress products and waste substances, and providing a healthy and efficient intracellular environment for the cell cycle process promoted by epigallocatechin gallate. The physical protective film formed by plant polysaccharides and hyaluronic acid, combined with the enhanced cell stress resistance and metabolic regulation ability of amino acids and trace elements, further consolidates the promoting effect of the plant component group on MSC proliferation and stemness maintenance. The composite growth factors are a mixture of BMP-2, TGF-β1, EGF, and IGF-1 in a mass ratio of (0.5-1.2):(0.3-0.8):(2-4):(3-8); the synergistic effect of multiple growth factors can activate the proliferation signaling pathway of MSCs and promote the cell cycle process; the natural plant extracts are a mixture of green tea polyphenols and rosmarinic acid in a mass ratio of 3:1, forming a composite defense system against oxidative stress. Green tea polyphenols provide immediate antioxidant protection, while rosmarinic acid achieves long-term defense through pathway regulation, forming a "scavenging-inhibiting" dual antioxidant mechanism. In addition, they maintain the stemness of MSCs through the synergistic effect of epigenetic and transcriptional regulation, improving the anti-interference ability of the culture system; in cooperation with hydroxypropyl-β-cyclodextrin, the chemical stability of the culture medium is enhanced.

[0025] (3)The culture medium additive for MSCs disclosed by the present invention constructs a complex regulatory network that mimics the in vivo microenvironment through the synergistic design of multiple components such as composite growth factors, plant component microemulsion, amino acids, and plant polysaccharides. This multi-target and multi-level intervention strategy breaks through the single-action mode of the prior art and represents a technological leap from "single-factor regulation" to "system microenvironment simulation", enabling MSCs to maintain their complete differentiation ability while proliferating at a high speed. Detailed implementation manners

[0026] The following description is used to disclose the present invention so that those skilled in the art can implement the present invention. The preferred embodiments described below are only examples, and those skilled in the art can think of other obvious variations.

[0027] Example 1

[0028] A culture medium additive for MSCs, comprising the following components: 5 μg / L of composite growth factors, 1 g / L of plant component microemulsion, 10 mg / L of amino acids, 0.3 g / L of plant polysaccharides, 0.1 g / L of hyaluronic acid, 100 mg / L of betaine, 40 mg / L of vitamin C, 0.6 mg / L of trace elements, 10 mg / L of natural plant extracts, 10 mg / L of hydroxypropyl-β-cyclodextrin, and 2 mmol / L of acetylglutamine; the composite growth factors are composed of BMP-2, TGF-β1, EGF, and IGF-1 mixed in a mass ratio of 0.5:0.3:2:3.

[0029] The preparation method of the plant component microemulsion includes the following steps: Dispersing curcumin, silybin, and epigallocatechin gallate in an organic solvent, adding an emulsifier, and mixing evenly to obtain a mixed solution; mixing the mixed solution with deionized water, and treating it 5 times under a pressure of 100 MPa by a high-pressure homogenizer to obtain the plant component microemulsion; the mass ratio of curcumin, silybin, epigallocatechin gallate, organic solvent, and emulsifier is 6:4:3:40:0.65; the organic solvent is absolute ethanol; the emulsifier is poloxamer 188; the volume ratio of the mixed solution to deionized water is 1:3.

[0030] The amino acid is proline; the plant polysaccharide is seaweed polysaccharide; the weight-average molecular weight of the hyaluronic acid is 100 kDa; the trace elements are composed of Se and Zn mixed in a mass ratio of 0.1:0.5; the natural plant extracts are composed of green tea polyphenols and rosmarinic acid mixed in a mass ratio of 3:1.

[0031] A preparation method of the culture medium additive for MSCs includes the following steps: Mixing each component, adding it to deionized water, stirring evenly under the conditions of 4 °C and darkness, then filling it into a freeze-drying bottle, pre-freezing at -60 °C for 2 hours, and then drying it in a freeze dryer for 25 h to obtain the culture medium additive for MSCs; the vacuum degree of the drying is 5 Pa, and the temperature is -40 °C.

[0032] An application of the culture medium additive for MSCs in MSC cell culture, adding the culture medium additive for MSCs to a basic cell culture medium for MSC cell culture; the addition amount of the culture medium additive for MSCs is 0.3 g / L; the basic cell culture medium is DMEM / F12 complete medium.

[0033] Example 2

[0034] A culture medium additive for MSCs, comprising the following components: 7 μg / L of composite growth factors, 1.5 g / L of plant component microemulsion, 13 mg / L of amino acids, 0.5 g / L of plant polysaccharides, 0.13 g / L of hyaluronic acid, 110 mg / L of betaine, 50 mg / L of vitamin C, 0.8 mg / L of trace elements, 15 mg / L of natural plant extracts, 15 mg / L of hydroxypropyl-β-cyclodextrin, and 2.5 mmol / L of acetylglutamine; the composite growth factors are a mixture of BMP-2, TGF-β1, EGF, and IGF-1 in a mass ratio of 0.7:0.5:2.5:4.

[0035] The preparation method of the plant component microemulsion includes the following steps: dispersing curcumin, silybin, and epigallocatechin gallate in an organic solvent, adding an emulsifier, and mixing evenly to obtain a mixed solution; mixing the mixed solution with deionized water, and treating it 6 times under a pressure of 110 MPa by a high-pressure homogenizer to obtain the plant component microemulsion; the mass ratio of curcumin, silybin, epigallocatechin gallate, organic solvent, and emulsifier is 6.5:4.5:3.5:45:0.8; the organic solvent is propylene glycol; the emulsifier is Tween80; the volume ratio of the mixed solution to deionized water is 1:3.

[0036] The amino acid is glycine; the plant polysaccharide is dextran; the weight-average molecular weight of the hyaluronic acid is 200 kDa; the trace elements are a mixture of Se and Zn in a mass ratio of 0.15:0.7; the natural plant extracts are a mixture of green tea polyphenols and rosmarinic acid in a mass ratio of 3:1.

[0037] A preparation method of the culture medium additive for MSCs includes the following steps: mixing each component, adding it to deionized water, stirring evenly under the conditions of 6 °C and in the dark, then loading it into a freeze-drying bottle, pre-freezing at -60 °C for 2 hours, and then drying it in a freeze dryer for 27 h to obtain the culture medium additive for MSCs; the vacuum degree of the drying is 6 Pa, and the temperature is -40 °C.

[0038] An application of the culture medium additive for MSCs in the culture of MSC cells, adding the culture medium additive for MSCs to a basic cell culture medium for MSC cell culture; the addition amount of the culture medium additive for MSCs is 0.3 g / L; the basic cell culture medium is DMEM / F12 complete medium.

[0039] Example 3

[0040] A culture medium additive for MSCs, comprising the following components: 9 μg / L of a composite growth factor, 2 g / L of a plant component microemulsion, 15 mg / L of amino acids, 1 g / L of plant polysaccharides, 0.15 g / L of hyaluronic acid, 130 mg / L of betaine, 60 mg / L of vitamin C, 0.9 mg / L of trace elements, 20 mg / L of a natural plant extract, 20 mg / L of hydroxypropyl-β-cyclodextrin, and 3 mmol / L of acetylglutamine; the composite growth factor is a mixture of BMP-2, TGF-β1, EGF, and IGF-1 in a mass ratio of 0.9:0.6:3:5.

[0041] The preparation method of the plant component microemulsion includes the following steps: dispersing curcumin, silybin, and epigallocatechin gallate in an organic solvent, adding an emulsifier, and mixing evenly to obtain a mixed solution; mixing the mixed solution with deionized water, and treating it 7 times under a pressure of 130 MPa by a high-pressure homogenizer to obtain the plant component microemulsion; the mass ratio of curcumin, silybin, epigallocatechin gallate, the organic solvent, and the emulsifier is 7:5:4:50:1.1; the organic solvent is absolute ethanol; the emulsifier is Tween80; the volume ratio of the mixed solution to deionized water is 1:3.

[0042] The amino acids are glutamic acid; the plant polysaccharides are seaweed polysaccharides; the weight-average molecular weight of the hyaluronic acid is 300 kDa; the trace elements are a mixture of Se and Zn in a mass ratio of 0.2:0.75; the natural plant extract is a mixture of green tea polyphenols and rosmarinic acid in a mass ratio of 3:1.

[0043] A preparation method of the culture medium additive for MSCs includes the following steps: mixing each component, adding it to deionized water, stirring evenly under the conditions of 7 °C and in the dark, then loading it into a freeze-drying bottle, pre-freezing at -60 °C for 2 hours, and then drying it in a freeze dryer for 28 h to obtain the culture medium additive for MSCs; the vacuum degree of the drying is 7.5 Pa, and the temperature is -40 °C.

[0044] An application of the culture medium additive for MSCs in MSC cell culture, adding the culture medium additive for MSCs to a basic cell culture medium for MSC cell culture; the addition amount of the culture medium additive for MSCs is 0.3 g / L; the basic cell culture medium is DMEM / F12 complete medium.

[0045] Example 4

[0046] A culture medium additive for MSCs, comprising the following components: 12 μg / L of composite growth factors, 2.5 g / L of plant ingredient microemulsion, 18 mg / L of amino acids, 1.3 g / L of plant polysaccharides, 0.18 g / L of hyaluronic acid, 140 mg / L of betaine, 75 mg / L of vitamin C, 1.1 mg / L of trace elements, 25 mg / L of natural plant extracts, 25 mg / L of hydroxypropyl-β-cyclodextrin, and 3.5 mmol / L of acetylglutamine; the composite growth factors are composed of BMP-2, TGF-β1, EGF, and IGF-1 mixed in a mass ratio of 1.1:0.7:3.5:7.

[0047] The preparation method of the plant ingredient microemulsion comprises the following steps: dispersing curcumin, silybin, and epigallocatechin gallate in an organic solvent, adding an emulsifier, and mixing evenly to obtain a mixed solution; mixing the mixed solution with deionized water, and treating it 7 times under a pressure of 140 MPa by a high-pressure homogenizer to obtain the plant ingredient microemulsion; the mass ratio of curcumin, silybin, epigallocatechin gallate, the organic solvent, and the emulsifier is 7.5:5.5:4.5:55:1.3; the organic solvent is composed of absolute ethanol and propylene glycol mixed in a mass ratio of 1:2; the emulsifier is composed of poloxamer 188 and Tween 80 mixed in a mass ratio of 3:5; the volume ratio of the mixed solution to deionized water is 1:3.

[0048] The amino acids are composed of proline, glycine, glutamic acid, and arginine mixed in a mass ratio of 1:1:2:3; the plant polysaccharides are composed of seaweed polysaccharides and dextran mixed in a mass ratio of 1:2; the weight-average molecular weight of the hyaluronic acid is 400 kDa; the trace elements are composed of Se and Zn mixed in a mass ratio of 0.25:0.9; the natural plant extracts are composed of green tea polyphenols and rosmarinic acid mixed in a mass ratio of 3:1.

[0049] A preparation method of the culture medium additive for MSCs comprises the following steps: mixing each component, adding it to deionized water, stirring evenly under the conditions of 9 °C and light avoidance, then filling it into a freeze-drying bottle, pre-freezing at -60 °C for 2 hours, and then drying it in a freeze dryer for 29 h to obtain the culture medium additive for MSCs; the vacuum degree of the drying is 9 Pa, and the temperature is -40 °C.

[0050] An application of the culture medium additive for MSCs in the culture of MSC cells, adding the culture medium additive for MSCs to a basic cell culture medium for MSC cell culture; the addition amount of the culture medium additive for MSCs is 0.3 g / L; the basic cell culture medium is DMEM / F12 complete medium.

[0051] Example 5

[0052] A culture medium additive for MSCs, comprising the following components: 13 μg / L of composite growth factors, 3 g / L of plant component microemulsion, 20 mg / L of amino acids, 1.5 g / L of plant polysaccharides, 0.2 g / L of hyaluronic acid, 150 mg / L of betaine, 80 mg / L of vitamin C, 1.2 mg / L of trace elements, 30 mg / L of natural plant extracts, 30 mg / L of hydroxypropyl-β-cyclodextrin, and 4 mmol / L of acetylglutamine; the composite growth factors are a mixture of BMP-2, TGF-β1, EGF, and IGF-1 in a mass ratio of 1.2:0.8:4:8.

[0053] The preparation method of the plant component microemulsion includes the following steps: dispersing curcumin, silybin, and epigallocatechin gallate in an organic solvent, adding an emulsifier, and mixing evenly to obtain a mixture; mixing the mixture with deionized water, and treating it 8 times under a pressure of 150 MPa by a high-pressure homogenizer to obtain the plant component microemulsion; the mass ratio of curcumin, silybin, epigallocatechin gallate, organic solvent, and emulsifier is 8:6:5:60:1.5; the organic solvent is absolute ethanol; the emulsifier is poloxamer 188; the volume ratio of the mixture to deionized water is 1:3.

[0054] The amino acid is arginine; the plant polysaccharide is seaweed polysaccharide; the weight-average molecular weight of the hyaluronic acid is 500 kDa; the trace elements are a mixture of Se and Zn in a mass ratio of 0.3:1; the natural plant extracts are a mixture of green tea polyphenols and rosmarinic acid in a mass ratio of 3:1.

[0055] A preparation method of the culture medium additive for MSCs includes the following steps: mixing each component, adding it to deionized water, stirring evenly under the conditions of 10 °C and in the dark, then filling it into a freeze-drying bottle, pre-freezing at -60 °C for 2 hours, and then drying it in a freeze dryer for 30 h to obtain the culture medium additive for MSCs; the vacuum degree of the drying is 10 Pa, and the temperature is -40 °C.

[0056] An application of the culture medium additive for MSCs in the culture of MSC cells, adding the culture medium additive for MSCs to a basic cell culture medium for the culture of MSC cells; the addition amount of the culture medium additive for MSCs is 0.3 g / L; the basic cell culture medium is DMEM / F12 complete medium.

[0057] Control Example 1 A culture medium additive for MSCs and its application are basically the same as those in Example 1, except that the plant component microemulsion is not added.

[0058] Control Example 2 A culture medium additive for MSCs and its application are basically the same as those in Example 1, except that hyaluronic acid and betaine are not added.

[0059] To further illustrate the beneficial technical effects of the culture medium additives for MSCs involved in the embodiments of the present invention, relevant performance tests were conducted on the culture medium additives for MSCs involved in Examples 1-5 and Comparative Examples 1-2. The test results are shown in Table 1, and the test methods are as follows: According to the application methods of each example, the cells were cultured in an incubator at 37°C and 5% CO2. The growth of the cells was observed every day, and the culture medium was changed every 3 days. After 7 days of culture, the cell viability was calculated by the trypan blue staining method. The CCK-8 method was used to detect the cell proliferation, and the proliferation rate of MSCs at 72 h was statistically analyzed; the MSCs used for the test were umbilical cord tissue-derived MSCs.

[0060] Table 1 Performance test results of the culture medium additives for MSCs Project Example 1 Example 2 Example 3 Example 4 Example 5 Comparative Example 1 Comparative Example 2 Cell viability (%) 98.5 99.0 99.3 99.5 99.9 96.2 97.0 Cell proliferation rate (%) 355.6 358.0 360.0 361.5 363.3 337.8 343.9 As can be seen from Table 1, the culture medium additives for MSCs involved in the embodiments of the present invention can significantly improve the viability and proliferation rate of MSCs. Compared with the comparative examples, the effects are significant; the combined use of plant component microemulsion, hyaluronic acid and betaine is beneficial to improving the above properties.

[0061] The above embodiments are only for illustrating the technical concept and characteristics of the present invention, and the purpose is to enable those skilled in the art to understand the content of the present invention and implement it accordingly. It should not be used to limit the protection scope of the present invention. All equivalent changes or modifications made according to the spirit and essence of the present invention should be covered within the protection scope of the present invention.

Claims

1. A culture medium additive for MSCs, characterized in that, It consists of the following components: composite growth factors 5 - 13 μg / L, plant ingredient microemulsion 1 - 3 g / L, amino acids 10 - 20 mg / L, plant polysaccharides 0.3 - 1.5 g / L, hyaluronic acid 0.1 - 0.2 g / L, betaine 100 - 150 mg / L, vitamin C 40 - 80 mg / L, trace elements 0.6 - 1.2 mg / L, natural plant extracts 10 - 30 mg / L, hydroxypropyl-β-cyclodextrin 10 - 30 mg / L, acetylglutamine 2 - 4 mmol / L; The composite growth factors are composed of BMP-2, TGF-β1, EGF, and IGF-1 mixed in a mass ratio of (0.5 - 1.2):(0.3 - 0.8):(2 - 4):(3 - 8); The preparation method of the plant ingredient microemulsion includes the following steps: dispersing curcumin, silybin, and epigallocatechin gallate in an organic solvent, adding an emulsifier, and mixing evenly to obtain a mixed solution; mixing the mixed solution with deionized water, and treating it 5 - 8 times under a pressure of 100 - 150 MPa by a high-pressure homogenizer to obtain the plant ingredient microemulsion; The mass ratio of curcumin, silybin, epigallocatechin gallate, organic solvent, and emulsifier is (6 - 8):(4 - 6):(3 - 5):(40 - 60):(0.65 - 1.5); The amino acids are at least one of proline, glycine, glutamic acid, and arginine; The plant polysaccharides are at least one of seaweed polysaccharides and dextran; The trace elements are composed of Se and Zn mixed in a mass ratio of (0.1 - 0.3):(0.5 - 1); The natural plant extracts are composed of green tea polyphenols and rosmarinic acid mixed in a mass ratio of 3:

1.

2. The culture medium additive for MSC according to claim 1, characterized in that, The organic solvent is at least one of absolute ethanol and propylene glycol; The emulsifier is at least one of poloxamer 188 and Tween 80; The volume ratio of the mixed solution to deionized water is 1:

3.

3. The culture medium additive for MSC according to claim 1, wherein The weight-average molecular weight of the hyaluronic acid is 100 - 500 kDa.

4. A method for preparing a culture medium additive for MSC according to any one of claims 1-3, characterized in that, It includes the following steps: mixing each component, adding it to deionized water, stirring evenly under the conditions of 4 - 10 °C and in the dark, then loading it into a freeze-drying bottle, pre-freezing at -60 °C for 2 hours, and then drying it in a freeze dryer for 25 - 30 h to obtain the MSC culture medium additive; The vacuum degree of the drying is 5 - 10 Pa, and the temperature is -40 °C.

5. Use of a culture medium additive for MSCs according to any one of claims 1 - 3 in MSC cell culture, characterized in that, Adding the MSC culture medium additive to the basic cell culture medium for MSC cell culture; The addition amount of the MSC culture medium additive is 0.1 - 0.5 g / L.

6. Use of the culture medium additive for MSC according to claim 5 in the culture of MSC cells, characterized in that, The basic cell culture medium is DMEM / F12 complete medium.