大鼠骨髓间充质干细胞向肝细胞的分化.docVIP

大鼠骨髓间充质干细胞向肝细胞的分化 徐洪军，关晓辉，关兴卓 Hepatocyte differentiation from rat marrow mesenchymal stem cells Xu Hong-jun, Guan Xiao-hui, Guan Xing-zhuo Abstract BACKGROUND: Mesenchymal stem cells (MSCs) can be induced into hepatocytes under microenvironment of hepatocyte growth factor in vitro. OBJECTIVE: To explore whether MSCs of adult rats can be induced into hepatocytes in vitro. DESIGN, TIME AND SETTING: The experiment was performed at Beihua University between October 2006 and May 2008. MATERIALS: Adult SD rats, irrespective of gender, weighing 180-230 g, were selected. METHODS: MSCs were collected from the femora of SD rats by Percoll density gradient centrifugation and. MSCs (1×108 L-1) were cultured in culture medium containing 20 μg/L hepatocyte growth factor. The culture medium was changed regularly. MAIN OUTCOME MEASURES: Flow cytometric analysis and alkaline phosphatase (AKP) staining were used for identification of cell surface marker and cell type. Morphological changes of MSCs were observed by phase-contrast microscope. On days 1, 3, 7, 14, 21 and 28, expressions of albumin, alpha fetoprotein (AFP) and CK18 of MSCs were also detected with immunohistochemistry technique. RESULTS: After culture, the volume of cell increased with a variety of forms, and adherent cells differentiated into large polygon and spindle-shaped cells with rich cytoplasm and multi-nuclei. Cell growth in early inoculation was faster than in advanced stage. MSCs collected from the femora of SD rats expressed antigens of CD29, CD44 and CD90, but not CD34 or CD45; AKP staining was negative for all MSCs. Cell purity was over 98%. Immunocytochemical analysis for CK18, albumin and AFP showed positive staining reaction for AFP on day 7 and for CK18 and albumin on day 14. CONCLUSION: MSCs of adult rats cultured with high concentration (20 μg/L) of hepatocyte growth factor can differentiate into hepatocytes. Xu HJ, Guan XH, Guan XZ.Hepatocyte differentiation from rat marrow mesenchymal ste