Maintenance of primary hepatocyte functions in vitro by inhibiting mechanical tension-induced YAP activation

It has been a long-standing challenge to maintain the functions of hepatocytes in vitro. In the present study, we found that mechanical tension-induced yes-associated protein (Yap) activation triggered hepatocyte dedifferentiation. Alleviation of mechanical tension by confinement of cell spreading was sufficient to inhibit hepatocyte dedifferentiation. Based on this finding, we identified a small molecular cocktail LBDXL through reiterative chemical screening that could maintain hepatocyte functions over the long term and in vivo repopulation capacity by targeting actin polymerization and actomyosin contraction. Hepatocytes mRNA profiles of freshly isolated hepatocytes (FH), freshly isolated Yap-/- hepatocytes (FH Yap-/-), LBDXL hepatocytes at three weeks (LBDXL 3W), BM hepatocytes at three weeks (BM 3W), BM Yap-/- hepatocytes at three weeks (BM 3W Yap-/-), BM hepatocytes at one day (BM 1D) and BM hepatocytes at three days (BM 3D) were generated by deep sequencing, in duplicate, using illumina Hiseq4000