| dc.description.abstract | Liver fibrosis is a life-threatening disease that significantly affects human life. Hepatic fibrosis is characterized by the production and deposition of connective tissue materials, which is mainly caused by the transformation of hepatic stellate cells (HSCs). In this study, HSCs isolation was established, and different glucose concentration (100mg/dL, 450mg/dL and 600mg/dL) in serum-free DMEM medium (supplement with 0.5mM L-Glutamine, 10 uM vitamin A and 50 ng/ml insulin) was investigated to stimulate the proliferation as well as maintain the quiescence. HSCs were examined the proliferation and activation at day 3, day 5, and day 7 of culture. The results showed that HSCs yield was 656.83 ± 182.29 (x103) cells per gram liver (n=5). These cells possessed specific morphology that has vitamin A/lipid droplet in the cytoplasm; positive with desmin marker. The isolated HSCs were successfully cultured in seven days showed the phenotypic change, the decrease of lipid droplets, and the expression of α-SMA. With the supplements of L-Glutamine, vitamin A and insulin without FBS, the 600mg/dL glucose medium induced a highest proliferation rate (31.85% , 21.82%, 13.19% cells in S and G2/M stage in day 3,5 and 7 respectively) on the HSCs but the HSCs decreased the lipid droplet storage compared to lower glucose concentration. The 100mg/dL glucose medium maintained the highest % lipid droplet area within HSC at three time points (16.17 ± 1.84, 12.69 ± 1.41, 9.64 ± 0.61 in day 3, 5, and 7 respectively).
Keywords: hepatic stellate cells, proliferation, activation, glucose, lipid droplets | en_US |
