Method for differentiating osteoplastic cells by oriented inducing chick embryo primordial germ cell

A technique for differentiation of primordial germ cells and osteoblasts, applied in the fields of therapeutic cloning, tissue engineering, and cell engineering, can solve the problems of few research reports on chicken EPGCs, and achieve the effects of strong repeatability, simple operation, and simple process

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 osteoplastic cells by oriented inducing chick embryo primordial germ cell

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Embodiment Construction

[0019] 1. Preparation of inducer

[0020] With 90% DMEM, 10% FBS, 5.5×10 -5 β-mercaptoethanol, 1×10 -7 mol / L dexamethasone, 0.01mol / L sodium β-glycerophosphate, and 0.05g / L vitamin C were used to prepare inducers.

[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-sugar DMEM culture medium. EPGCs were cultured for 5 ...

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Abstract

The invention discloses the method used to directionally induce chick embryo primordial germ cell to differentiate into bone cell in vitro. It uses the inducer formed by 90% DMEM, 10% FBS, 5.5*10-5 beta-mercaptoethanol, 1*10-7mol/L dexamethasone, 0.01mol/L beta-sodium glycerophosphate, 0.05g/L vitamin C to process induction differentiating culturing in vitro for chick embryo primordial germ to differentiate into bone cell finally. It can build cell model for clinical medicine study, widely apply in tissue engineering field for providing reliable experience data, supply new source for cell substituting therapy and tissue therapy, even organ transplantation. The invention has the advantages of simple technology, convenient operation, strong repeatability, no need for special apparatus, only need to master universal cell separating and culturing operation.

Description

technical field [0001] The invention relates to a method for obtaining osteoblasts through directional induction and differentiation culture of chicken embryo primordial germ cells in vitro, and belongs to the technical fields of therapeutic cloning, cell engineering and tissue engineering. Background technique [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. Martin et al. successfully induced osteoblasts from bone marrow stromal cells in vitro; Holtor et al. studied the role of dexamethasone in the process of induction and differentiation of mouse bone marrow mesenchymal stem cells into osteoblasts, and proved that in the absenc...

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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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