[Cultivation and karyotype analysis of the human embryonic stem cells HUES4].
OBJECTIVE
To establish a protocol for culturing of human embryonic stem cells (HUES4) without any animal-derived feeder cells and to investigate the karyotype stabilities of HUES4 cells after long-term cultivation.


METHODS
HUES4 cells were cultured on mitomycin C treated MEFs or human foreskin fibroblast feeder cells. The pluripotency of the ES cells was analyzed by immunocytochemistry staining to detect the expression of pluripotent marker, karyotype of the ES cells at passage 27, 34, 41, 44 and short tandem repeat (STR) at passage 27 were analyzed.


RESULTS
The HUES4 cells cultured on human feeder cells were positive for alkaline phosphatase activity, SSEA-4, TRA-1-60 and TRA-1-81 staining, but negative for SSEA-1. Analysis of karyotype at different passages suggested an abnormal karyotype 46, XY, t(9;15)(q22;q26) mosaicism occurred in HUES4, and the ratios of abnormal increased with passage.


CONCLUSIONS
HUES4 could be cultured without animal-derived feeder cells and the incidence of abnormal karyotype might be increased with long-term culture.
Zhang
Wen-hong
WH
Obstetrics & Gynecology Research Institute, Third Affiliated Hospital, Guangzhou Medical College, Guangzhou, Guangdong, 510150 PR China.
Sun
Xiao-fang
XF
Kong
Shu
S
Jiang
Yong-hua
YH
Zheng
Yu-hong
YH
Yin
Yi-fei
YF
D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D049109:Cell Proliferation; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D052816:Foreskin / Q000166:cytology; D006801:Humans; D007150:Immunohistochemistry; D007621:Karyotyping / Q000379:methods*; D008297:Male; D018895:Microsatellite Repeats / Q000235:genetics; D011110:Polymorphism, Genetic