Fibroblast Induced By Hepa1-6Promotes The Proliferation And Differenation Of Mouse Liver Stem Cells

Objective: To investigate how the supernate of myofibroblast influencethe proliferation and differentiation of fetal mouse liver stem cells HP14.5.Methods: The Hepa1-6supernate of mouse liver cancer cells was usedto activate fetal mouse fibroblasts extracted and collected ordinarysupernate of fibroblast and the supernate of myofibroblast to prepare thesupernate of fibroblast conditioned culture medium (CMFb) and thesupernate of myofibroblast conditioned medium (CMFAb). The HP14.5liver stem cells were divided in three group to develop: The DMEM culturemedium treatment group (DMEM group), CMFb culture medium treatmentgroup (CMFb group) and CMFAb culture medium treatment group(CMAFb). the MTT assay was used to draw cell growth curves for thethree groups, the immunofluorescence staining and real-time quantitativepolymerase chain reaction to detect the albumin (ALB), alpha-fetoprotein(AFP), cytokeratin18(CK18) in protein and mRNA expression level,and the PAS staining method to detect Glycogen synthesis condition.Results: The MTT assay showed faster proliferation of liver stem cellsin CMAFb group, compared with other groups; The immunofluorescencestaining and Real-Time PCR showed that after5days of HP14.5cultured,the level of the ALB, albumen in CK18, mRNA expression and Glycogensynthesis in CMAFb group was much higher than those in CMFb group and DMEM group, while the number alpha-fetoprotein and mRNAexpression level declined remarkably.Conclusion: The fibroblasts activated by Hepa1-6is able to increasethe proliferation of liver stem cells and differentiate the functional matureliver cells.