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Method for isolated culture of human fat mesenchyma stem cell and special culture medium thereof

A technique for isolating and culturing stromal cells, which is applied in the field of isolating and culturing human adipose-derived mesenchymal stem cells, and achieves the effect of simple and high-efficiency cultivation methods.

Active Publication Date: 2008-12-03
微能生命科技集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in practical application, it faces ethical challenges or limitations of materials

Method used

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  • Method for isolated culture of human fat mesenchyma stem cell and special culture medium thereof
  • Method for isolated culture of human fat mesenchyma stem cell and special culture medium thereof
  • Method for isolated culture of human fat mesenchyma stem cell and special culture medium thereof

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Embodiment 1

[0020] Example 1. Isolation and culture of human adipose-derived mesenchymal stem cells

[0021] 1. Reagents

[0022] Collagenase type I was purchased from Sigma. Trypsin was purchased from Gibco. DMEM / F12 (ie DF12), M199, MEM, RPMI1640, BME, IMEM and DMEM were purchased from GIBCO Company. MCDB-201 was purchased from Sigma Company. EGF was purchased from Gibco. PDGF, VEGF and bFGF were purchased from Sigma Company. Hydrocortisone was purchased from Sigma. FCS and horse serum (HS) were purchased from Hyclone Company.

[0023] 2. Isolation and culture of human adipose-derived mesenchymal stem cells

[0024] The following three media were used for separation and culture: A. The final concentration was 580ml / L DMEM / F12, 400ml / L MCDB-201, 20ml / L fetal bovine serum (FCS), 10ng / ml EGF, 10ng / ml PDGF composition.

[0025] B. The final concentration is 580ml / L DMEM / F12, 419ml / L MCDB-201, 1ml / L fetal bovine serum (FCS), 1ng / ml EGF, 1ng / ml PDGF.

[0026] C. The final concentrat...

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Abstract

The invention discloses a method for separately culturing a human adipose mesenchymal stem cell and a dedicated culture medium thereof. The culture medium used for separately culturing the human adipose mesenchymal stem cell comprises an animal cell basic culture medium, fetal calf serum, an epidermal growth factor and a platelet-derived growth factor. The final concentration of the fetal calf serum is 1-200 mL / L, the final concentration of the epidermal growth factor is 1-100 ng / ml, and the final concentration of the platelet-derived growth factor is 1-100 ng / ml. The adipose mesenchymal stem cell of the invention has CD31-, CD34-, CD45- and HLA-DR-, as well as the phenotype of CD29+, CD44+, CD105+ and Flk-1+. The specificity cell surface marker and the relevant antihelion molecule of a skeletal muscle cell and a vascular endothelia cell can be expressed after inducement is performed in vitro. Muscle fiber, vascular endothelin and functional muscle satellite cells can be differentiated in a muscle injury model mouse body caused by medicine and the expression of dystrophin protein on the ducheme muscular dystrophy (DMD) model mouse (mdx) myolemma can be partially recovered, so as to release the pathological symptom of the model mouse.

Description

technical field [0001] The invention relates to a method for separating and culturing human adipose mesenchymal stem cells and a special culture medium thereof. Background technique [0002] Stem cells are primitive cells with self-renewal ability and multi-lineage differentiation potential, and are ideal cells for cell transplantation. During individual development, there are various forms of stem cells, such as embryonic stem cells, pluripotent stem cells, and committed stem cells in various tissues such as hematopoietic stem cells, neural stem cells, etc. However, in practical application, it faces ethical challenges or limitations of material selection. Mesenchymal stem cells (MSC) are a subpluripotent stem cell population with multi-lineage differentiation potential in human tissues, because they can be induced to differentiate into a variety of tissue cells in a specific environment, and have low immunogenicity. It can form stable chimerism and other characteristics,...

Claims

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Application Information

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IPC IPC(8): C12N5/08C12N5/0775
Inventor 赵春华刘艳宁闫曦
Owner 微能生命科技集团有限公司
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