|
|
2022, Vol. 26 ›› Issue (16): 2510-2515Gelatin collagen composite hydrogel and inducible factor regulate differentiation of rat bone marrow mesenchymal stem cells into hepatocyte-like cellsYuan Yihang, Xu Menghan, Niu Xufeng
Abstract: BACKGROUND: In vitro artificial liver system has become a very effective and practical treatment for liver failure, but there are still some problems such as limited source of cells and limited function of cells. OBJECTIVE: To optimize and screen gelatin collagen composite hydrogel that meet the needs of cell growth, the hydrogel was used as the basal cell growth material to explore the scheme of inducing hepatic differentiation of bone marrow mesenchymal stem cells. METHODS: Gelatin collagen composite hydrogel with gelatin concentration of 50, 100, 150, 200 g/L was prepared by mixing gelatin solution and collagen solution at different volume ratios (1:1, 2:1, 4:1). The volume ratio of the solution and the gelatin concentration were selected for subsequent experiments by melting point, microstructure, water absorption and porosity. Rat bone marrow mesenchymal stem cells seeded in composite hydrogel were used as experimental group; rat bone marrow mesenchymal stem cells cultured alone were used as controls. Cell proliferation was detected by CCK-8 assay. The two-step induction regimen was used to induce the hepatic differentiation of two groups of rat bone marrow mesenchymal stem cells. The levels of albumin production and urea secretion in the supernatant were tested at 3, 7, 14, and 21 days after induced culture. RESULTS AND CONCLUSION: (1) After testing the melting point, microscopic morphology, water absorption and porosity of the experimental groups with different ratios, the composite gel was most suitable for cell culture when the 150 g/L gelatin solution was mixed with collagen solution at 4:1 volume ratio. (2) CCK-8 assay showed that the proliferation activity of the experimental group was lower than that of the control group at 1, 3 and 5 days, and the difference between the two groups gradually decreased, and there was no difference at 10 days. (3) Experiment of hepatic differentiation showed that with the extension of induction time, the amount of albumin secretion in the two groups increased gradually, and the amount of albumin in the experimental group was higher than that in the control group at 14 and 21 days (P < 0.05). The amount of urea secretion in the two groups increased significantly within 3-7 days of induction, and stabilized after 7 days. The amount of urea secretion in the experimental group was slightly higher than that in the control group at 14 and 21 days, but the difference was not significant (P > 0.05). (4) The results showed that gelatin collagen composite hydrogel could induce rat bone marrow mesenchymal stem cells to differentiate into hepatocyte-like cells. Key words:composite hydrogel, artificial liver, bone marrow mesenchymal stem cells, hepatocytes, induced differentiation, gelatin, collagen, EDC/NHS |