Establishment of human embryonic stem cell line from gamete donors.
BACKGROUND
Human embryonic stem (HES) cell derived from human blastocyst can be propagated indefinitely in the primitive undifferentiated state while remaining pluripotent. It has exciting potential in human developmental biology, drug discovery, and transplantation medicine. But there are insufficient HES cell lines for further study.


METHODS
Three oocyte donors were studied, and 3 in vitro fertilization (IVF) cycles were carried out to get blastocysts for the establishment of HES cell line. Isolated from blastocysts immunosurgically, inner cell mass (ICM) was cultured and propagated on mouse embryonic fibroblasts (MEFs). Once established, morphology, cell surface markers, karyotype and differentiating ability of the cell line were thoroughly analyzed.


RESULTS
Four ICMs from 7 blastocysts were cultured on MEFs. After culture, one cell line (cHES-1) was established and met the criteria for defining human pluripotent stem cells including a series of markers used to identify pluripotent stem cells, morphological similarity to primate embryonic stem cells and HES reported else where. Normal and stable karyotype maintained over 60 passages, and demonstrated ability to differentiate into a wide variety of cell types.


CONCLUSIONS
HES cell lines can be established from gamete donors at a relatively highly efficient rate. The establishment will exert a widespread impact on biomedical research.
Li
Tao
T
Reproductive Medical Center, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, China.
Zhou
Can-quan
CQ
Mai
Qing-yun
QY
Zhuang
Guang-lun
GL
D001755:Blastocyst / Q000166:cytology*; D002454:Cell Differentiation; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000032:analysis; D005260:Female; D005307:Fertilization in Vitro; D006801:Humans; D007621:Karyotyping; D008297:Male; D050814:Octamer Transcription Factor-3; D013234:Stem Cells / Q000166:cytology*; D014019:Tissue Donors; D014157:Transcription Factors / Q000032:analysis
D004268:DNA-Binding Proteins; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D014157:Transcription Factors