Derivation and characterization of human embryonic germ cells: serum-free culture and differentiation potential.
This study examined the effects of a chemically defined culture medium supplement, knock-out serum replacement (KSR), on the growth and differentiation of human embryonic germ cells (hEgc) and found that the efficiency of the initial establishment of hEGC lines in KSR medium was significantly higher than in fetal calf serum (FCS) medium. The percentage of undifferentiated hEGC colonies growing in KSR medium was significantly higher than in FCS-based medium (P < 0.05). The hEGC colonies showed typical mouse embryonic germ cell-like morphology. They showed normal and stable diploid karyotype and expressed alkaline phosphatase (AP), stage-specific embryonic antigens (SSEA) and other specific markers of pluripotent cells. In addition, hEGC could form simple and cystic embryoid bodies (EB) that consisted of various cell types including neural, epithelial and rhythmically beating cardiac cells, even sperm-like and oocyte-like cells. Tumour-like outgrowths were formed in nude mice and found to contain a variety of cell types, including uterine epithelium, adipocytes, squamous tissue and skin structures. In conclusion, an appropriate serum-free culture system has been developed for the establishment of hEGC lines. This may provide an in-vitro model to study differentiation and can be used as a potential source of therapy for infertility and regenerative medicine.
Hua
Jinlian
J
College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering and Technology, Shaanxi Key Lab for Agriculture Molecular Biotechnology Centre, Northwest A and F University, Yangling, Shaanxi, 712100 China. jlhua2003@126.com
Yu
Haisheng
H
Liu
Sheng
S
Dou
Zhongying
Z
Sun
Yadong
Y
Jing
Xiaoqi
X
Yang
Chunrong
C
Lei
Anmin
A
Wang
Huayan
H
Gao
Zhimin
Z
D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation; D002478:Cells, Cultured; D003470:Culture Media; D016895:Culture Media, Serum-Free; D017931:DNA Primers; D005854:Germ Cells / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D007621:Karyotyping; D051379:Mice; D008855:Microscopy, Electron, Scanning; D020133:Reverse Transcriptase Polymerase Chain Reaction
D003470:Culture Media; D016895:Culture Media, Serum-Free; D017931:DNA Primers