CHEN Shizuan,YU Fuxiang,CHEN Junyu, et al. Isolation and culture of hepatocytes and sinusoidal endothelial cells and preparation of hepatocyte scaffolds[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2019, 49(9): 630-633.
Abstract：Objective: To explore the methods of isolation and culture of rat hepatocytes and sinusoidal endothelial cells and preparation of hepatic acellular scaffolds. Methods: Rat hepatocytes isolated from liver tissues were digested by collagenase IV and purified in culture medium after washing, filtration and centrifugation. Cell immunohistochemistry was used to detect the cells after culture. Hepatic sinusoidal endothelial cells were isolated by Percoll density gradient centrifugation and purified in culture medium before they were detected by immunofluorescence. The scaffolds of rat hepatocyte acellular scaffolds were prepared by circulating perfusion and acellular treatment. The scaffolds were confirmed by scanning electron microscopy. Results: The isolated rat hepatocytes were round, full and clear under microscope. Immunohistochemical staining of cytokeratin 18 in hepatocytes revealed brown granules in the cytoplasm. Thehepatic sinusoidal endothelial cells were spindle-shaped and the nucleus was located in the center. Endothelin-1 immunofluorescence staining showed red signal on the cell surface. The scanning electron microscopy of the rat liver decellularized scaffold showed that the extracellular matrix was retained, and no nuclear and cytoplasmic components were found. Conclusion: Hepatocytes and sinusoidal endothelial cells are successfully isolated and hepatic acellular scaffolds were prepared.
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