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Serum-free human amniotic mesenchymal stem cell culture medium and preparation method thereof

A technology of mesenchymal stem cells and human amniotic membrane, applied in the field of stem cells, can solve the problems of unsatisfactory cell proliferation rate and poor cell adhesion, achieve good cell differentiation potential, simple preparation method, and enhance the effect of adhesion performance

Inactive Publication Date: 2015-09-02
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to solve the problems of poor cell adhesion and unsatisfactory cell proliferation rate in the prior art, the inventor obtained a new serum-free culture medium for human amniotic mesenchymal stem cells through a large number of tests and screenings. Culture flask coated, easy to operate, good cell adhesion and morphology, high cell proliferation rate

Method used

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  • Serum-free human amniotic mesenchymal stem cell culture medium and preparation method thereof
  • Serum-free human amniotic mesenchymal stem cell culture medium and preparation method thereof
  • Serum-free human amniotic mesenchymal stem cell culture medium and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Effect of Human Amniotic Mesenchymal Stem Cell Culture Medium with Different Formulas on Cell Proliferation

[0035] The inventors conducted a large number of test screenings on the serum-free human amniotic mesenchymal stem cell culture medium. The P1 generation of HAMSC at 5000 / cm 2 The density was inoculated in a six-well plate, culture medium 1, medium 4 and medium 5 were added respectively, the cells were digested with 0.25% trypsin every 3 days, the cells were collected and the number was calculated, and the number was calculated at 5000 / cm 2 The density was re-inoculated and cultured until the P5 generation, and the cell proliferation curve was drawn according to the number of cells harvested in each generation, and the results were as follows: figure 1 shown.

[0036] From figure 1 It can be seen that adding a certain concentration of putrescine hydrochloride and recombinant human activin A at the same time can greatly increase the cell proliferatio...

Embodiment 2

[0037] Example 2 Serum-free culture medium for human amniotic mesenchymal stem cells

[0038] Serum-free human amniotic mesenchymal stem cell culture medium, including IMDM basal medium, also includes the following components and their concentrations: recombinant human insulin 1mg / mL, recombinant transferrin 2.5μg / mL, vitamin C 16.1μg / mL, Human serum albumin 1mg / mL, bFGF 10ng / mL, PDGF-bb 10ng / mL, EGF 10ng / mL, IGF-1 10ng / mL, TGF-β10ng / mL, fibronectin 10ng / mL, fetuin 1mg / mL mL, putrescine hydrochloride 10 μg / mL and recombinant human activin A 10 ng / mL.

[0039] Preparation method: Add recombinant human insulin mother liquor, recombinant transferrin mother liquor, vitamin C mother liquor, human serum albumin, bFGF mother liquor, PDGF-bb mother liquor, EGF mother liquor, IGF-1 mother liquor to the IMDM basal medium according to the stated concentration , TGF-β mother liquor, fibronectin mother liquor, fetuin mother liquor, putrescine hydrochloride mother liquor and recombinant ...

Embodiment 3

[0041] Example 3 Serum-free culture medium for human amniotic mesenchymal stem cells

[0042] Serum-free human amniotic mesenchymal stem cell culture medium, including IMDM basal medium, also includes the following components and their concentrations: recombinant human insulin 1mg / mL, recombinant transferrin 2μg / mL, vitamin C 20μg / mL, human serum Albumin 1mg / mL, bFGF 15ng / mL, PDGF-bb 10ng / mL, EGF 10ng / mL, IGF-1 10ng / mL, TGF-β10ng / mL, fibronectin 15ng / mL, fetuin 3mg / mL, Putrescine hydrochloride 15 μg / mL and recombinant human activin A 10 ng / mL.

[0043] Preparation method: similar to the preparation method in Example 2.

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Abstract

The invention provides a serum-free human amniotic mesenchymal stem cell culture medium, and belongs to the technical field of stem cells. The serum-free human amniotic mesenchymal stem cell culture medium comprises the following components: an IMDM basal culture medium, recombinant human insulin, recombinant transferrin, vitamin C, human serum albumin, fibroblast growth factors, platelet-derived growth factors, epidermal growth factors, insulin-like growth factors, transforming growth factors, fibronectin, fetuin, putrescine salt and recombinant human activin. The serum-free human amniotic mesenchymal stem cell culture medium provided by the invention does not need a culture bottle coating process, and is good in cell adherence and morphology and high in cell proliferation rate.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and relates to a serum-free culture medium for human amniotic mesenchymal stem cells and a preparation method thereof. Background technique [0002] Human amniotic mesenchymal stem cells are derived from placental amniotic membrane tissue. They are a kind of adult stem cells with obvious plasticity and multi-lineage differentiation potential. Under the regulation of different growth factors, they can differentiate into different tissue cell types derived from three germ layers, such as Adipocytes, osteoblasts, liver-like cells, cardiomyocyte-like cells, glial cells, neuron cells and islet-like cells, etc. In addition, human amniotic mesenchymal stem cells do not express major histocompatibility class II antigens, but express a small amount of major histocompatibility class I antigens, have low immunogenicity, have little transplant rejection, and have immunoregulatory effects. The trophic fac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 陈海佳王一飞葛啸虎戴国胜冯德龙
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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