Human mesenchymal stem cell serum-free medium and preparation method thereof

A serum-free medium and stem cell technology, which is applied in the field of human mesenchymal stem cell serum-free medium and its preparation, and achieves the effects of good cell uniformity, clear components, and test results up to standard

Inactive Publication Date: 2018-07-06
XIAN TISSUE ENG & REGENERATIVE MEDICINE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the deficiencies in the existing serum-free culture of human mesenchymal stem cells

Method used

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  • Human mesenchymal stem cell serum-free medium and preparation method thereof
  • Human mesenchymal stem cell serum-free medium and preparation method thereof
  • Human mesenchymal stem cell serum-free medium and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0045] In order to solve the deficiencies in the existing serum-free culture of human mesenchymal stem cells; the present invention provides a serum-free medium for human mesenchymal stem cells, the main components of which include basal medium and additives.

[0046] The above basal medium is any one of a-MEM, L-DMEM, or DMEM-F12; the actual operation can be selected according to the needs of the specific cultured mesenchymal stem cells.

[0047]The above additives include human serum albumin (HSA), insulin-transferrin-selenium supplement (ITS), hydrocortisone, vitamin C (Vc), linoleic acid, progesterone, sodium pyruvate, non-essential amino acids, Essential amino acids, basic fibroblast growth factor (bFGF), transforming growth factor (TGF-β). Among them, HSA provides binding proteins for cell growth; ITS is a compound substance that promotes the use of glucose and amino acids by cells and also provides iron and selenium for cell growth; hydrocortisone can stimulate cell pro...

Embodiment 2

[0077] Taking human umbilical cord mesenchymal stem cells as an example, the culture medium of Example 1 is used for culturing, and the basal medium of the culture medium is a-MEM; the ratio of various additives in the medium is as follows: the amount of HSA added The added amount of ITS is 250μg / L, the added amount of ITS is 6ml / L, the added amount of vitamin C is 30mg / L, the added amount of hydrocortisone is 50μg / L, the added amount of linoleic acid is 2mg / L, and the added amount of sodium pyruvate The addition amount is 100mg / L, the addition amount of progesterone is 15μg / L, the addition amount of non-essential amino acid is 10ml / L, the addition amount of essential amino acid is 10ml / L, the addition amount of bFGF is 10μg / L, TGF-β The added amount is 10μg / L. This formula is mainly suitable for subculture of umbilical cord mesenchymal stem cells. Stem cells are easily aged or differentiated during the subculture process. Higher doses of vitamin C and progesterone are added t...

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Abstract

The present invention relates to a human mesenchymal stem cell serum-free medium, the main components of the human mesenchymal stem cell serum-free medium include a basal medium and an additive; the mesenchymal stem cell serum-free medium has a clear composition and no animal-derived component. A mesenchymal stem cell product produced by the human mesenchymal stem cell serum-free medium is used clinically without causing anaphylactic reaction; and the serum-free medium is simple in preparation method, does not require any special equipment, and is easy to be mass-produced; and mesenchymal stemcells produced by the human mesenchymal stem cell serum-free medium have good cell homogeneity and high proliferation rate. After repeated passages, detection results of cell surface markers are up to standard, and the cells still have good capability of differentiation into osteoblasts, adipocytes and chondrocytes.

Description

technical field [0001] The invention belongs to the technical field of mesenchymal stem cell culture, and in particular relates to a human mesenchymal stem cell serum-free medium and a preparation method thereof. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of adult stem cells that exist in bone marrow, fat, placenta, connective tissue and organ interstitium, and have the potential of self-renewal, proliferation and multidirectional differentiation, and can differentiate into osteoblasts. , chondrocytes, adipocytes, liver parenchymal cells, muscle cells, and neuron cells have become one of the important seed cells in the research fields of basic medicine, clinical medicine, regenerative medicine, and tissue engineering, and have very attractive application prospects. [0003] The traditional medium for culturing mesenchymal stem cells generally consists of basal medium and fetal bovine serum (FBS). FBS contains growth factors, hormones, carrier pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0662C12N5/0668C12N2500/25C12N2500/30C12N2500/32C12N2500/36C12N2500/38C12N2500/90C12N2501/115C12N2501/15C12N2501/39C12N2501/392C12N2501/998
Inventor 刘文佳程蕊苹邓志宏张永和
Owner XIAN TISSUE ENG & REGENERATIVE MEDICINE RES INST
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