The phenotypic characteristics of human fetal liver progenitors and clonal culture in vitro / 中华肝脏病杂志

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the phenotypic characteristics of human fetal liver cells (FLCs) and to obtain the homogenous hepatic progenitors with cloning.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>The clonal culture system is beneficial to obtain the homogenous hepatic progenitor cells from the heterogeneous culture of FLCs.</p>