[In-vitro amplification of oval cells with preservation of stem cell phenotype].
OBJECTIVE
To explore cell culture techniques for amplification of oval cells with preservation simultaneously of the stem cell characteristics.


METHODS
Oval cell line OC3 was cultured in RPMI 1640 supplemented with 15% fetal bovine serum and 20 µg/L EGF. Cells were harvested every 5 passages and were examined with biomarkers including OV-6, c-kit, gamma-glutamyl transpeptidase, placental form of glutathione-S-transferase (GST-P), pyruvate kinase M₂, pyruvate kinase L and albumin using techniques including RT-PCR, immunocytochemistry, and enzymo-cytochemistry.


RESULTS
OC3 cell lines could be amplified abundantly in-vitro associating with expression of infant liver cell markers at various level, including OV-6, c-kit, gamma-glutamyl transpeptidase, GST-P, pyruvate kinase M₂, but no expression of mature hepatocyte markers detected including pyruvate kinase L and albumin.


CONCLUSIONS
Amplification of OC3 cells with preservation of the stem cell phenotype and high proliferation index can be achieved up to the 79(th) passages by culturing in RPMI 1640 supplemented with 15% fetal bovine serum and 20 µg/L EGF.
Department of Pathology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
Qiong-rong
QR
Chen
Fang
F
Liu
Guo-qiang
GQ
Zhao
Ling
L
Xue
Rui-de
RD
Hu
Hui-xi
HX
Wu
Meng
M
Zhang
D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D003470:Culture Media; D005982:Glutathione Transferase / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D008099:Liver / Q000166:cytology* / Q000254:growth & development; D010641:Phenotype; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D011770:Pyruvate Kinase / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
D000943:Antigens, Differentiation; D003470:Culture Media; C115011:oval cell marker OV-6; D005982:Glutathione Transferase; D011770:Pyruvate Kinase; D019009:Proto-Oncogene Proteins c-kit