Serum-free adipose-derived stem cell culture medium and preparation method thereof

A technology of fat stem cells and culture medium, applied in the field of stem cells, can solve the problems of poor cell adhesion and unsatisfactory cell proliferation rate, and achieve the effect of simple preparation method, good cell differentiation potential, and promotion of growth and proliferation

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

AI Technical Summary

Problems solved by technology

[0006] 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 medium for adipose-derived stem cells through a large number of tests and screenings. This medium does not need to be coated with culture bottles. Simple, good cell adhesion and morphology, high cell proliferation rate

Method used

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  • Serum-free adipose-derived stem cell culture medium and preparation method thereof
  • Serum-free adipose-derived stem cell culture medium and preparation method thereof
  • Serum-free adipose-derived stem cell culture medium and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Effect of media containing different concentrations of stem cell growth factors on cell proliferation

[0033] The inventor conducted a large number of tests to screen and obtain the serum-free fat stem cell culture medium formula provided by the present invention, and further investigated the influence of the concentration of stem cell growth factors. The P1 generation of ADSCs at 5000 / cm 2The density was inoculated in six-well plates, and cultured with serum-free adipose-derived stem cell culture medium containing different concentrations of stem cell growth factors (compared with medium 1, the difference was only the concentration of stem cell growth factors was different), and the cells were divided every 3 days. Digest with 0.25% trypsin, collect the cells and calculate the number, and use 5000 cells / 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 harve...

Embodiment 2

[0035] Example 2 Serum-free adipose stem cell culture medium

[0036] Serum-free fat 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, polytea Phenol 10μg / mL and stem cell growth factor 10ng / mL.

[0037] 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 solution, fibronectin mother solution, fetuin mother solution, tea polyphenol mother solution and stem cell growth factor mother solution, stir evenly, pass through the membrane ...

Embodiment 3

[0039] Example 3 Serum-free adipose stem cell culture medium

[0040] Serum-free fat stem cell culture medium, including IMDM basal medium, also includes the following components and their concentrations: recombinant human insulin 2mg / mL, recombinant transferrin 2μg / mL, vitamin C 20μg / mL, human serum albumin 1mg / mL mL, bFGF 15ng / mL, PDGF-bb 10ng / mL, EGF 15ng / mL, IGF-1 10ng / mL, TGF-β10ng / mL, fibronectin 15ng / mL, fetuin 2mg / mL, tea polyphenol 15μg / mL and stem cell growth factor 15ng / mL.

[0041] Preparation method: similar to the preparation method in Example 2.

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Abstract

The invention provides a serum-free adipose-derived stem cell culture medium, belonging to the technical field of stem cells. The serum-free adipose-derived stem cell culture medium comprises an IMDM (Iscove's modified Dulbecco's medium) basal culture medium, recombinant human insulin, recombinant transferrin, vitamin C, human serum albumin, fibroblast growth factor, platelet-derived growth factor, epidermal growth factor, insuline-like growth factor, transforming growth factor, fiber connexin, fetuin, tea polyphenol and stem cell growth factor. The serum-free adipose-derived stem cell culture medium provided by the invention does not need to perform culture bottle coating, and has the advantages of cell adherent property, favorable morphology and high cell proliferation rate.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and relates to a serum-free fat stem cell culture medium and a preparation method thereof. Background technique [0002] Adipose-derived stem cells (ADSCs) are a kind of stem cells isolated from adipose tissue with multi-directional differentiation potential. Studies have found that ADSCs can proliferate stably in vitro and have a low death rate. At the same time, they have a wide range of sources, a large reserve in the body, and are easy to obtain. A large number of stem cells can be obtained from a small amount of tissue. They are suitable for large-scale culture and have little damage to the body. [0003] The traditional stem cell medium containing animal serum brings many unfavorable factors to the production and scientific research of stem cells. Serum can provide stem cells with hormones, growth factors, transfer proteins and other nutrients needed for growth and proliferation, but it...

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王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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