Umbilical cord mesenchymal stem cell serum-free medium and preparation method thereof

Abstract

The invention relates to the technical field of stem cell culture, and relates to a serum-free medium for umbilical cord mesenchymal stem cells and a preparation method thereof, including DMEM low-sugar medium, human serum albumin, transferrin, epidermal growth factor, transforming growth factor , insulin, olive oil, danshensu sodium and ginkgolic acid. The culture medium prepared by the present invention can significantly increase the proliferation quantity and proliferation rate of umbilical cord mesenchymal stem cells, and umbilical cord mesenchymal stem cells are not easy to differentiate to produce phenotypic changes, better protect umbilical cord mesenchymal stem cells, and umbilical cord mesenchymal stem cells The effect of sticking to the wall is good, the cost is low, and it is easy to process production.

Description

technical field [0001] The invention relates to the technical field of stem cell culture, in particular to a serum-free medium for umbilical cord mesenchymal stem cells and a preparation method thereof. Background technique [0002] Umbilical cord mesenchymal stem cells refer to a kind of multifunctional stem cells existing in neonatal umbilical cord tissue, which can differentiate into many kinds of tissue cells. Human umbilical cord mesenchymal stem cells can be successfully expanded by using human umbilical cord serum as the main culture system. The growth of umbilical cord mesenchymal stem cells depends on the existence of serum, such as fetal bovine serum or newborn bovine serum. In the medium without adding serum, most cells (including stem cells) cannot grow, proliferate or differentiate, and serum can provide multiple Nutrients necessary for the growth of many cells (including stem cells), such as insulin, adrenal cortex hormones, steroid hormones (estradiol, testost...

AI Technical Summary

Problems solved by technology

[0003] Serum is a very complex mixture, and there may be as many as hundreds of components. At present, its exact composition, content and mechanism of action are still unclear. The quality of serum varies greatly between batches, which is affected by the source of raw materials of the manufacturer. It is very difficult to ensure the consistency of each batch, which is not conducive to the standardization of laboratory research or product production. The serum may contain some unknown or undetected harmful microorganisms such as mycoplasma and viruses, which may cause potential damage to cells. In addition, some substances such as antibodies, complements and bacterial toxins in the serum also have adverse effects on the growth of cells, and even cause cell death.

[0004] However, the clinical application of stem cells requires the cultivation of serum-free medium, and the cultivation effect of serum-free medium on the market is far inferior to that of serum medium. Chinese patent application CN105420182A discloses a serum-free umbilical cord mesenchymal stem cell culture medium. Including DMEM medium, basic fibroblast growth factor, epidermal growth factor, insulin, leukemia inhibitory factor, trypsin, 2-mercaptoethanol, sodium selenite and fibronectin, etc., achieved by adding a higher content of fibronectin Good adhesion performance, but the proliferation rate of cells is slow; Chinese patent application CN102634482A discloses a serum-free complete medium for mesenchymal stem cells, which is composed of basal medium and added components, and the basal medium is DMEM / F12, the added ingredients are composed of recombinant insulin, human serum albumin, human transferrin, cholesterol, sodium selenite, ferric nitrate and dextran, which do not contain serum and avoid the serum containing serum medium There are defects such as instability between batches, cytotoxicity, and a large number of heterologous proteins, but the cultured stem cells are easy to differentiate and cause phenotypic changes. Disadvantages such as unsatisfactory wall performance

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