Hepatocytes ; cytology ; Liver, Artificial

Hepatocytes ; cytology ; Humans ; Liver ; cytology ; Stem Cells ; cytology

Cell Death ; Hepatocytes ; cytology ; pathology ; Humans

Hepatic Stellate Cells ; cytology ; Hepatocytes ; pathology

Animals ; Hepatocytes ; cytology ; Mice ; Stem Cell Transplantation ; Stem Cells ; cytology

Animals ; Endothelial Cells ; cytology ; Hepatocytes ; cytology ; Humans ; Liver Regeneration

OBJECTIVETo investigate the phenotypic characteristics of human fetal liver cells (FLCs) and to obtain the homogenous hepatic progenitors with cloning.

METHODSImmunofluorescence 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.

RESULTSThe 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.

CONCLUSIONSThe clonal culture system is beneficial to obtain the homogenous hepatic progenitor cells from the heterogeneous culture of FLCs.

Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Fetus ; cytology ; Hepatocytes ; cytology ; Humans ; Stem Cells ; cytology

OBJECTIVETo investigate the possibility of transdifferentiation of adipose mesenchymal stem cells (AMSCs) into hepatocytes.

METHODSHuman omentum adipose tissue was dispersed with collagenase I. Cells collected were cultured in a DMEM-F12 medium containing 2% FBS supplemented with 20 ng/ml HGF, 10 ng/ml FGF4, 1xITS and 0.1 micromol/L dexasmison. The cells of the control group were also cultured in the same kind of medium but without any cytokines serving as a control. The expression of hepatic transcriptional factors such as GATA4 and HNF1 were checked by RT-PCR. At the end of the induction, hepatocyte markers were analysed by flow cytometry, and cytokeratin expressions were examined using cyto-immunofluorescence methods.

RESULTSAMSCs grew like fibroblasts and were passaged easily. Most of the third passaged AMSCs were positive against anti-CD29, anti-CD44 antibodies, but negative for the anti-CD34 and anti-CD45 ones. The hepatic transcriptional factor was expressed gradually to higher levels during the induction time. AFP and Alb positive cells were 30.0% and 17.8% of the total cultured cells, and the rate of cells positive to the two markers was 6.9%. The inducted cells were positive for CK18 and CK19 antibodies at the end of the induction. The cells in the control group were negative when checked by these methods.

CONCLUSIONSAMSCs could be directed to differentiate into hepatocytes in vitro by a cytokine cocktail with a low concentration FBS culture system.

Adipocytes ; cytology ; Cell Differentiation ; Cell Transdifferentiation ; Cells, Cultured ; Hepatocytes ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology

OBJECTIVETo investigate the biological function of hepatocyte-like cells derived from mesenchymal stem cells that isolated from human umbilical cord UC-MSCs in vitro, and to detect the changes in the immunogenicity of the differentiated hepatocyte-like cells (DHC).

METHODSTransdifferentiation of UC-MSCs into hepatic lineage in vitro was induced in modified two-step induction medium. The expressions of hepatic specific markers were detected by RT-PCR analysis and immunofluorescence staining at different time points after induction. The levels of albumin and urea in the supernatants of cultures were measured by enzyme-linked immunosorbent assay. Furthermore, the immunosuppressive property of DHC was detected by one-way mixed lymphocyte culture.

RESULTSThe mRNA and proteins of alpha fetoprotein (AFP), albumin (ALB),and cytokeratin-19 (CK-19) were expressed in naive UC-MSCs at low levels. DHC highly expressed hepatic markers AFP, ALB, CK-19, and tryptophan 2, 3-dioxygenase 14 and 28 days after hepatic differentiation and were accompanied by an increased production of ALB and urea in supernatant in a time-dependent manner. DHC did not express human leukocyte antigen DR antigen and significantly decreased the lymphocyte proliferation.

CONCLUSIONUC-MSCs are able to differentiate into functional hepatocyte-like cells in vitro, while the immunogenicity of DHC remains low.

Cell Transdifferentiation ; Cells, Cultured ; Hepatocytes ; cytology ; immunology ; Humans ; Mesenchymal Stromal Cells ; cytology ; Umbilical Cord ; cytology

Hepatocytes ; cytology ; Humans ; Liver Failure ; therapy ; Liver, Artificial