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Factors influencing the efficiency of isolation and culture of human embryonic germ cells

Published online by Cambridge University Press:  12 February 2007

Hua Jin-Lian ,
Dou Zhong-Ying ,
Xu Xiao-Ming ,
Li Song ,
Yang Yu-Ai  and
Lei An-Min
Hua Jin-Lian
Affiliation:
Shaanxi Centre of Stem Cells Engineering & Technology, Northwest Sci-Tech University of Agriculture & Forestry, Yangling 712100, China
Dou Zhong-Ying*
Affiliation:
Shaanxi Centre of Stem Cells Engineering & Technology, Northwest Sci-Tech University of Agriculture & Forestry, Yangling 712100, China
Xu Xiao-Ming
Affiliation:
Shaanxi Centre of Stem Cells Engineering & Technology, Northwest Sci-Tech University of Agriculture & Forestry, Yangling 712100, China
Li Song
Affiliation:
Shaanxi Centre of Stem Cells Engineering & Technology, Northwest Sci-Tech University of Agriculture & Forestry, Yangling 712100, China
Yang Yu-Ai
Affiliation:
Shaanxi Centre of Stem Cells Engineering & Technology, Northwest Sci-Tech University of Agriculture & Forestry, Yangling 712100, China
Lei An-Min
Affiliation:
Shaanxi Centre of Stem Cells Engineering & Technology, Northwest Sci-Tech University of Agriculture & Forestry, Yangling 712100, China
*
*Corresponding author: E-mail: [email protected]
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Abstract

Embryonic germ (EG) cells are pluripotent cells derived from the primordial germ cells of gonads, gonadal ridges and mesenteries, and analogies of foetuses, with the ability to undergo both self-renewal and multiple differentiation. These cells can differentiate into derivatives of all three embryonic germ layers when transferred to an in vitro environment and have the ability to form any fully differentiated cell of the body. The present paper investigates some factors influencing the efficiency of isolation and culture of human EG cells, such as foetus age, culture serum, added cytokines and feeder cells. The results demonstrate that foetuses of 7–12 weeks are optimal for in vitro culture of human EG cells. The basic medium consisted of DMEM, 1×non-essential amino acids, 2 mM l-glutamine and 1 mM sodium pyruvate. Supplementation with 15% foetal bovine serum, 4 ng/ml human recombinant leukaemia inhibitory factor, 4 ng/ml basic fibroblast growth factor and 20 ng/ml stem cell factor clearly improved the efficiency of isolation and culture of human EG cells. Murine embryonic fibroblasts were better feeder cells than human embryonic fibroblasts, bovine embryonic fibroblasts or STO cell line.

Keywords

humansembryonic germ cellsembryonic stem cellscytokinesfeeder cells
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 1 , Issue 3 , December 2004 , pp. 191 - 196
Copyright
Copyright © China Agricultural University and Cambridge University Press 2004

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