Human embryonic germ cells isolation from early stages of post-implantation embryos.
Human embryonic germ (hEG) cell is a very important alternative pluripotent stem cell resource. We describe the derivation of hEG cells from human embryonic fetal gonads over 6-8 weeks postconception. A large number of EG-like cell clumps were obtained at passage 1 and thus facilitated the following routine culture when the donor tissues were trypsinized with gentle pipetting and plated on feeder layer cells in the initial culture. Eight diploid hEG cell lines have been cultivated in vitro for extended periods while maintaining expression of markers characteristic of pluripotent stem cells. Human EG cells expressed transcription factor Oct4, a marker of pluripotency in mouse EG cells, at a high and steady level. Expression of markers indicative of differentiation along the three germ lineages was also observed in EBs. High level of alkaline phosphatase activity was shown in EG cells. These encouraging findings provide a starting point for potential applicability of hEG cells.
Liu
ShanRong
S
Department of Embryology and Histology, Second Military Medical University, Xiang'yin Road 800, 200433, Shanghai, People's Republic of China.
Liu
HouQi
H
Pan
YuQiong
Y
Tang
ShuPing
S
Xiong
Jun
J
Hui
Ning
N
Wang
ShuLing
S
Qi
ZhongTian
Z
Li
LingSong
L
D000469:Alkaline Phosphatase / Q000378:metabolism; D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology* / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D005854:Germ Cells / Q000166:cytology*; D005865:Gestational Age; D006801:Humans; D007621:Karyotyping; D050814:Octamer Transcription Factor-3; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D015415:Biomarkers; D004268:DNA-Binding Proteins; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D012333:RNA, Messenger; D014157:Transcription Factors; D000469:Alkaline Phosphatase