Hepatocyte-like cells differentiation of human amniotic fluid colony-derived stem cells using Transwell 3-D co-culture

Abstract

Objective To investigate the potential of human amniotic fluid colony-derived stem cells(h AFCSCs) differentiated into hepatocyte-like cells using transwell co-culture. Methods Using colony poculum, h AFCSCs were isolated from second-trimester amniotic fluid.After cultured and amplified, the expression of surface antigens was identified by flow cytometry.The h AFCSCs were induced into hepatocyte-like cells using transwell co-culture.Observe the change of cell morphology after co-culture. The expression of alpha-fetoprotein(AFP), Albumin(ALB), Cytokeratin-18(CK-18)and hepatocyte gene was detected with immunofluorescence and reverse transcriptase-polymerase chain reaction(RTPCR). Results h AFCSCs were positive for CD44, CD90, and negative for CD10, CD14, CD34, CD45 and human leukocyte antigen-DR(HLA-DR).After 2-4 weeks induction, hepatocyte-like cells appeared.It confirmed that these cells expressed AFP, ALB, CK-18, GATA 4, Hepatocyte nuclear factor (HNF)-1αand cytochrome 20A(CYP20A). Conclusion Human amniotic fluid colony-derived stem cells can be differentiated into hepatocyte-like cells using transwell co-culture. Key words: Human amniotic fluid colony-derived stem cells; Transwell; 3-D co-culture; Hepatocyte-like cells