Results: Results1. Primary liver perfusion combined with density gradient centrifuge via 60% percoll can obtain multiple quantity of HSCs up to 1.8 × 107 per rat, and cell viability ≥90%. 2. Initial target cells adhered to cultural dish presented rutond. Gradually, target cells presented typical star-like cells after culture 7 days. It was observed that “wreath” lipid around the nucleus, quenched green fluorescence excited by 328 nm, positive for nile red and HSCs specific markers (Desmin, GFAP), the cell purity ≥95%. 3. Primary HSCs expressed stem cell markers such as P75NTR, CD90, CD105, Dlk-1, JQ1 c-kit,

CD133, sox2, nanog, oct-4, lgr5 in mRNA level. Moreover, A-769662 concentration P75NTR, CD90, c-kit, sox-2 and

oct-4 have found expression in protein.4. Compared to control group, the morphology of primary HSCs has changed after induction by bFGF 20 ug/L, FGF4 20 ug/L, HGF 20 ug/L, IL-6 1 ug/L that became rotund or polygonal.5. Compared to control group, the hepatocyte specific markers of primary HSCs has expressed after induction by bFGF 20 ug/L, FGF4 20 ug/L, HGF 20 ug/L, IL-6 1 ug/L that both in mRNA transcription of AFP (p < 0.05) and ALB (p < 0.01) and protein level.6. Compared to control group, the stem cell markers of primary HSCs has dramatically decreased after induction by bFGF 20 ug/L, FGF4 20 ug/L, HGF 20 ug/L, IL-6 1 ug/L in the mRNA transcription of P75NTR (p < 0.05) and CD90, c-kit, sox-2, oct-4 (p < 0.01). Proteins for stem cell markers were also significantly decreased click here or even undetectable. Conclusion: Conclusion1. The study reported an available method acquiring for high quality and quantity of primary HSCs of rat liver.2. Primary

HSCs expressed stem cell markers such as P75NTR, CD90, c-kit, sox-2 and oct-4, in which both mRNA transcription and protein level, which speculated HSCs might possess stem cell characteristics.3. HSCs had probability of differentiation to hepatocyte-like cells by cytokines induction in vitro. Meanwhile, HSCs expressed hepatocyte specific marker AFP and ALB both in gene and protein.4. During induction by cytokines, HSCs’ stem cell markers has decreased significantly, which possibly prompted HSCs might have potential stem cell characteristics and a group of potential stem cells or progenitors in liver5. The study provided a method of cytokines induction to differentiate HSCs to hepatocyte-like cells in a short time, which were the foundation of further mechanism study.6. Stem cell markers of P75NTR, CD90, c-kit, sox2 and oct-4 might be available in further study on HSCs stem cell characteristics. Key Word(s): 1. stellate cells; 2. stem cell; 3. induction; 4.