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Method for differentiating fat cells by oriented inducing chick embryo primordial germcell

A technology for differentiation of primordial germ cells and adipocytes, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problems of few research reports on chicken EPGCs, achieve strong repeatability, scientific and reasonable preparation, The effect of easy operation

Inactive Publication Date: 2007-03-28
YANGZHOU UNIV
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Problems solved by technology

At present, as an important embryonic stem cell, most studies on its directed differentiation are found in bone marrow mesenchymal stem cells, mouse embryonic stem cells, and bone marrow stromal cells, while there are few reports on chicken EPGCs.

Method used

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  • Method for differentiating fat cells by oriented inducing chick embryo primordial germcell

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Embodiment Construction

[0018] 1. Preparation of inducer

[0019] ①Inducer I: 90%DMEM+10%FBS+5.5×10 -5 β-mercaptoethanol+1×10 -6 mol / L dexamethasone+10mg / L insulin+5×10 -4 mol / L 3-isobutyl-1-methylxanthine.

[0020] ②Inducer II: 90%DMEM+10%FBS+5.5×10 -5 β-mercaptoethanol + 10mg / L insulin.

[0021] 2. Isolation, culture and passage of chicken embryo primordial germ cells

[0022] The EPGCs aggregated in the gonads were extracted at the 19th and 28th stages of chicken embryo development by a separate enzyme dissociation method, and the extracted cells were inoculated on the prepared feeder layer, and added with 10% fetal bovine serum, 2% chicken serum, 2mmol / L L-glutamine, 1mmol / L sodium pyruvate, 5.5×10 -5 mol / L β-mercaptoethanol, 10μl / ml non-essential amino acids, 10IU / ml murine leukemia inhibitory factor (mLIF), 5ng / ml human stem cell growth factor (hSCF), 10ng / ml basic fibroblast growth factor (bFGF), 0.04ng / ml human interleukin-11 (hIL-11), 10ng / ml insulin-like growth factor (hIGF) in high-...

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Abstract

The invention relates to the method used to directionally induce chick embryo primordial germ cell to differentiate into fat cell. The adopted inducer includes inducer I and II which are respectively formed by 90% DMEM, 10% FBS, 5.5*10-5 beta-mercaptoethanol, 1*10-6mol / L dexamethasone, 10mg / L insulin, 5*10-4mol / L 3-isobutyl-1-methylxanthine, and 90% DMEM, 10% FBS, 5.5*10-5 beta-mercaptoethanol, 10mg / L. The invention has the advantages of simple technology, scientific and reasonable inducer and basic medium, convenient operation, strong repeatability. And it can provide new source for cell substituting therapy and tissue therapy, even organ transplantation.

Description

technical field [0001] The invention relates to a method for directionally inducing the differentiation of chicken embryo primordial germ cells into adipocytes, and belongs to the technical fields of therapeutic cloning, cell engineering and tissue engineering. technical background [0002] Embryonic Primordial germ cells (EPGCs) are cells in the most primitive form of the embryonic germ line and are the progenitor cells of sperm or eggs. As a kind of pluripotent stem cells, EPGCs, like ES cells, can differentiate into other types of cells under appropriate conditions in vitro, and are one of the most promising seed cells in clinical medicine and tissue engineering research. Jiang et al. successfully induced adult bone marrow mesenchymal stem cells into adipocytes, and Yagami induced mesenchymal stem cells isolated from human cartilage, and the results showed that mesenchymal stem cells differentiated into adipocytes. At present, as an important embryonic stem cell, most st...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/077
Inventor 李碧春葛剑辉陈国宏秦洁吴信生赵文明徐琪孙国波戴建明孙鹏翔魏彩霞
Owner YANGZHOU UNIV
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