SFM (serum-free medium) for culturing MSCs (mesenchymal stem cells)

Abstract

The invention relates to an SFM (serum-free medium) for culturing MSCs (mesenchymal stem cells). Based on volume, the SFM comprises the following components: 10.2 grams per liter of alpha-MEM (alpha-minimum essential medium), 2.4 grams per liter of sodium bicarbonate, 1 to 5 millimoles of L-glutamine, 50 to 300 milligrams per liter of poloxamer 188, 2 to 8 grams per liter of recombinant human albumin, 10 to 20 milligrams per liter of recombinant human transferrin, 2 to 10 milligrams per liter of recombinant human insulin, 1 to 5 millimoles per liter of Hepes, 50 nanomoles of beta-mercaptoethanol, 0.1 to 1 milligram per liter of lipid, 1 to 5 milligrams per liter of trace element, 0.1 to 5 milligrams per liter of glutathione, 0.5 to 5 milligrams per liter of para-aminobenzoic acid, 1 to 50 nanograms per milliliter of hydrocortisone, 20 to 50 milligrams per liter of vitamin PP, 5 to 50 milligrams per liter of vitamin C, 2 to 10mu M of compound shown in a formula I, 5 to 20mu M of compound shown in a formula II, 10 to 20 nanograms per milliliter of progestin, 1 to 10 milligrams per liter of putrescine, 1 to 10 international units per liter of heparin, 1 to 10 nanograms per milliliter of EGF (epidermal growth factor), 1 to 10 nanograms per milliliter of b-FGF (b-fibroblast growth factor), 1 to 10 nanograms per milliliter of HGF (hepatocyte growth factor) and 1 to 10 nanograms per milliliter of VEGF (vascular endothelial growth factor). The SFM for culturing the MSCs is a BPS-SFM which has determinate chemical components and is free of animal-derived substances.

Description

Technical field [0001] The invention relates to a serum-free medium for culturing mesenchymal stem cells, belonging to the technical field of cell engineering and biomedicine. Background technique [0002] Mesenchymal stem cells (MSCs) are adult stem cells that are widely present in human tissues and organs. Because of their high self-renewal ability and multi-directional differentiation potential, they are used in clinical hematopoietic support, promotion of stem cell implantation, and immune regulation. It has potential application prospects. At present, large-scale preclinical and clinical studies have confirmed that MSCs are directly infused as cell therapy products or combined with scaffold materials to construct tissue-engineered tissues and organs (skin, bone or cartilage, etc.) for transplantation. It has good safety and effectiveness in the treatment of burns, bone defects, soft tissue filling, end-stage liver disease, myocardial infarction, diabetes and other trauma or...

AI Technical Summary

Problems solved by technology

[0007] (3) High batch-to-batch variability makes it difficult to standardize the cell production process;

[0008] (4) Serum may also contain growth inhibitory factors and cytotoxic substances;

[0009] (5) It is very difficult to carry out quality control to reduce various pollution risks;

[0010] (6) The effects of other unknown factors, hormones and other substances are yet to be studied

Serum-free medium has the characteristics of clear components, and can well maintain the proliferation and differentiation pluripotency of MSCs in vitro, but there are problems such as high price and short storage time

At the same time, most of these media need auxiliary gelatin and other matrigel to coat the culture plate, which not only may introduce animal-derived components, but also increases the workload of cell culture expansion and the chance of contamination during passage

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