Clinical-grade serum-free culture medium for stem cells

Abstract

The invention discloses a clinical-grade serum-free culture medium for stem cells. According to the culture medium, an L-DMEM culture medium, an IMDM culture medium, a DMEM-F12 or alpha-MEM culture medium, an mTeSRTM1 culture medium, an E8 culture medium, a KSR conditioned culture medium and the like are used as basic culture media, and lipid substances such as inositol and linoleic acid and synthetic lipid substance components are removed; and meanwhile the culture medium contains the following added components: polyvinyl alcohol, bone morphogenetic protein BMP-4, vitamin C, retinoic acid, nicotinamide and azithromycin. The culture medium is clear in composition, controllable in quality and free of animal serum components, can be used for culturing various stem cells including pluripotent stem cells and multipotent stem cells, and can maintain the capability of stemness of the stem cells without differentiation after multiple passages.

Description

technical field [0001] The invention relates to a medium for cell culture, in particular to a serum-free medium for clinical-grade stem cells, which belongs to the field of biomedicine. Background technique [0002] The current basal medium cannot support the long-term culture of stem cells, and there is still room for optimization. Now, more research is looking at the effects of supplementation on stem cells. A certain amount of fetal bovine serum is usually added to the traditional stem cell culture process, but fetal bovine serum has several disadvantages: high cost, batch variation, limited availability, serum is susceptible to contamination by viruses, mycoplasma and other pathogens, and heteroimmunity The possibility of reaction, at the same time, serum contains a large number of amino acids, nucleosides, proteins, hormones, lipids and other trace components, the content and specific role of these components are still not fully determined, the lipids contained in seru...

AI Technical Summary

Problems solved by technology

In the traditional stem cell culture process, a certain amount of fetal bovine serum is usually added, but fetal bovine serum has several disadvantages: high cost, batch variation, limited availability, serum is susceptible to contamination by viruses, mycoplasma and other pathogens, and heteroimmunity The possibility of reaction, at the same time, serum contains a large number of amino acids, nucleosides, proteins, hormones, lipids and other trace components, the content and specific role of these components are still not fully determined, the lipids contained in serum can promote cell differentiation

In order to avoid the above defects in the use of fetal bovine serum, a medium that can replace animal serum has been developed. The most commonly used ones are human AB serum, human platelet lysate and serum-free chemically defined medium, but each has its own shortcomings. AB serum Both human platelet lysate and human platelet lysate have the possibility of virus contamination, and there are obvious differences between different batches; various growth factors released by human platelet lysate may have substances that inhibit cell growth, and change the expression of cell surface markers and Cell function, etc. Although the current chemically defined medium does not contain serum and can avoid potential risks brought by serum products, it is not perfect. It can promote stem cell differentiation and improve immunogenicity, but cannot maintain the stemness of stem cells well.

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