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A serum-free medium for mesenchymal stem cells

A technology of serum-free medium and mesenchymal stem cells, which is applied in the field of serum-free medium for mesenchymal stem cells, and can solve the problems that the content of ingredients and specific functions are not completely determined.

Active Publication Date: 2019-09-13
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Serum contains a large number of trace components such as amino acids, nucleosides, proteins, hormones, and lipids. The content and specific effects of these components have not yet been fully determined

Method used

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  • A serum-free medium for mesenchymal stem cells
  • A serum-free medium for mesenchymal stem cells
  • A serum-free medium for mesenchymal stem cells

Examples

Experimental program
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Effect test

Embodiment 1

[0015] A serum-free medium for mesenchymal stem cells in this embodiment uses L-DMEM medium as the base medium, and also includes the following additional components, and the additional components and their contents (final concentrations) are as follows: 25 μg / mL of laminin, laminin Adhesin 25 μg / mL, transferrin 0.2 ng / mL, trypsin 10 μg / mL, aprotinin 0.1 mg / mL, growth hormone 20 ng / mL, insulin 1 U / mL, hydrocortisone 0.5 μg / mL, dexamethasone Methasone 0.2nM, bFGF 20ng / mL, EGF 30ng / mL, B27 volume fraction 1%, IGF-I 10ng / mL, IGF-II 30ng / mL, choline 30nM, linoleic acid 10nM, sodium selenite 30nM, phosphoric acid Ethanolamine 50nM, glutathione 2.5μg / mL, vitamin C 50μg / mL, vitamin E 30μg / mL, vitamin B12 10μM, biotin 20μM.

[0016] The specific preparation method is to add the above-mentioned supplementary components to the L-DMEM medium, and make the final concentration to be the above-mentioned concentration, thereby obtaining the serum-free medium for mesenchymal stem cells.

Embodiment 2

[0018] A serum-free medium for mesenchymal stem cells in this embodiment is based on DMEM-F12 medium, and also includes the following additional components, and the additional components and their contents (final concentrations) are as follows: laminin 40 μg / mL, Transferrin 2ng / mL, trypsin 20μg / mL, aprotinin 0.1mg / mL, growth hormone 30ng / mL, insulin 10U / mL, hydrocortisone 1μg / mL, dexamethasone 0.3nM, bFGF 40ng / mL , EGF 20ng / mL, B27 volume fraction 2%, IGF-I 10ng / mL, IGF-II 20ng / mL, choline 30nM, linoleic acid 20nM, sodium selenite 15nM, phosphoethanolamine 30nM, glutathione 5μg / mL, vitamin C 50 μg / mL, vitamin E 30 μg / mL, vitamin B12 20 μM, biotin 30 μM.

[0019] Its specific preparation method is to add the above-mentioned additional components to the DMEM-F12 medium, and make the final concentration to the above-mentioned concentration, thereby obtaining the serum-free medium for mesenchymal stem cells.

Embodiment 3

[0021] A serum-free medium for mesenchymal stem cells in this embodiment uses α-MEM medium as the base medium, and also includes the following additional components, and the additional components and their contents (final concentrations) are as follows: laminin 50 μg / mL, Transferrin 0.2ng / mL, trypsin 30μg / mL, aprotinin 0.1mg / mL, growth hormone 30ng / mL, insulin 6U / mL, hydrocortisone 1μg / mL, dexamethasone 0.3nM, bFGF 30ng / mL mL, EGF 30ng / mL, B27 volume fraction 3%, IGF-I 10ng / mL, IGF-II10ng / mL, choline 20nM, linoleic acid 20nM, sodium selenite 20nM, phosphoethanolamine 20nM, glutathione 2.5 μg / mL, Vitamin C 50μg / mL, Vitamin E 30μg / mL, Vitamin B12 20μM, Biotin 30μM.

[0022] The specific preparation method is to add the above-mentioned supplementary components to the α-MEM medium, and make the final concentration to be the above-mentioned concentration, thereby obtaining the serum-free medium for mesenchymal stem cells.

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Abstract

The invention discloses a mesenchymal stem cell serum-free culture medium. The culture medium takes an L-DMEM, DMEM-F12 or alpha-MEM culture medium as a basic culture medium and also includes the following addition components: fibronectin, laminin, transferrin, trypsin, aprotinin, growth hormone, insulin, hydrocortisone, dexamethasone, bFGF, EGF, B27, IGF-I, IGF-II, choline, linoleic acid, sodium selenite, phosphorylethanolamine, glutathione, vitamin C, vitamin E, vitamin B12 and biotin. The culture medium has the advantages of defined composition and controllable quality, and mesenchymal stem cells cultured by the culture medium grow normal; and after multiple passages, the 'dryness' of the mesenchymal stem cells and the ability of the mesenchymal stem cells to be differentiated into osteoblasts, chondrocytes, adipocytes, neurons and other types of cells are still kept.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a serum-free medium for mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (MSCs) are pluripotent stem cells derived from mesoderm with high self-renewal ability and multi-directional differentiation potential. It is isolated and can be differentiated into osteoblasts, chondrocytes, adipocytes and neurons under specific conditions. With a wide range of sources and no ethical and moral issues, MSC has been proven in many ways to be an ideal seed cell for autologous and allogeneic cell transplantation and treatment. At present, a number of clinical application studies of MSCs have been carried out at home and abroad, involving various types of MSCs from various sources and various types of diseases. [0003] The medium components used in the preparation of clinically applied stem cell preparations should have sufficient purity and meet the quality ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775C12N5/02
Inventor 李志远程娜王飞林佐贤叶威
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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