[The phenotypic characteristics of human fetal liver progenitors and clonal culture in vitro].
OBJECTIVE
To investigate the phenotypic characteristics of human fetal liver cells (FLCs) and to obtain the homogenous hepatic progenitors with cloning.


METHODS
Immunofluorescence and flow cytometry were used to determine the phenotypes of the FLCs. The proliferating colonies were isolated using clone ring in different culture conditions. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to determine the mRNA expression after further cultivation.


RESULTS
The cultured FLCs showed a non-typical epithelial morphology. The positive rate for hepatic cell specific markers alpha-fetoprotein (AFP), albumin (Alb), cytokeratin 8 (CK8) and CK19 were approximately 28.1%, 84.7%, 55.1% and 9.1% respectively. Furthermore, the FLCs expressed the hematopoietic stem cell markers CD34 and CD45 with percentages of 0.04% and 0.09%. 71.8% and 75.3% of the FLCs were positive for the mesenchymal cell marker CD105 and CD166. Most of the colonies showed an elongated morphology, some with an unregular spreading-out morphology, only a small number of colonies with an epithelial-like morphology. RT-PCR results showed that among the 19 colonies obtained after further cultivation and the percentages of epithelial cell adhesion molecule (EpCAM), AFP, Alb and CK19 were 52.6%, 21.1%, 52.6% and 84.2%, respectively.


CONCLUSIONS
The clonal culture system is beneficial to obtain the homogenous hepatic progenitor cells from the heterogeneous culture of FLCs.
Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China.
Ping
P
Wang
Wei-hong
WH
Li
Bao-qing
BQ
Liu
Xin
X
Lu
Hong
H
You
Ji-dong
JD
Jia
Hai-yan
HY
Zhang
D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D005333:Fetus / Q000166:cytology*; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D013234:Stem Cells / Q000166:cytology*