Effect of Soft Fibrin Gels onTransdifferentiation Behaviour in Activated Hepatic Stellate Cell / 中山大学学报(医学科学版)

ABSTRACT

[Objective]To determine whether activated hepatic stellate cell(HSC)become quiescent phenotype if cultured in a soft extracellular matrix(ECM).[Methods]HSC-T6 cells which stably expressed myofibroblast(MFB) phenotype,were cultured in the 2D and 3D in-vitro matrix culture models that were constructed by 1 mg/mL fibrin gel. The proliferation activity of HSC-T6 was determined by cell counting kit-8(CCK8)assay at different time points(24 h, 48 h,72 h,96 h). Immunofluorescence and gelatin zymography analysis were performed to detect the expression of α-SMA,MMP-2,and MMP-9. Moreover,the cellular surface morphology and deformability was detected by AFM.[Results]After cultured in the fibrin gel,the cellular proliferation activity,expression of α-SMA,MMP-2 and MMP-9,and cellular deformability of activated HSC-T6 was obviously down-regulated in contrast to the control group (P<0.05),and there were also differences in the inhibition of cellular proliferation activity,the assembly of α-SMA stress fiber and the effect of cellular deformability between 2D and 3D fibrin gel cultured(P<0.05).[Conclusions]Soft fibrin gel inhibited the transdifferentiation of activated HSC-T6,but could not make it revert to α-SMA negative quies-cent phenotype.