Differentiation characteristics of rat mesenchymal stem cells in vitro

    KANG Xinª²Qin, ZANG Weiª²Jin, XU Xiaoª²Li, SONG Tuª²Sheng, Yu Xiaoª²Jiang, ZENG Juª²Rong

    1Department of Biology, 2Division of Cardiovascular Physiology & Pharmacology, College of Medicine, Xi¬ðan Jiaotong University, Xi¬ðan 710061, China

    ¡¾Abstract¡¿ AIM: To study the differentiating characteristics of rat mesenchymal stem cells in vitro. METHODS: The rat marrow stem cells were isolated by Percoll (1.073 kg/L). The third passage cells had been induced by 0.1 ¦Ìmol/L dexamethasome, 10 mmol/L ¦Âª²glycerophosphate and 50 ¦Ìmol/L ascorbic acidª²2ª²phosphate. On the 7th day and 14th day, the cells were stained by modified Gomori to show the activities of alkaline phosphatase. On the 21st day, the cells were stained by Von Kassa to show calcium salts. The MSCs were induced by 1 ¦Ìmol/L Dexamethasome, 10 mg/L bovine insulin, 0.25 mmol/L 1 methylª²3ª²isobutylxamthine and 100 ¦Ìmol/L indomethacin. The MSCs were induced by 5 mmol/L ¦Âª²mercaptoethanol for 5 h and analyzed by immunohistochemistry. RESULTS: At the 7th and 14th day after osteoblastic induction, 11.5% and 37.5% of the cells stained with modified Gomori were positive, respectively. At the 21st day, the calcium salts were observed by Von Kassa. After 3 and 7 days¬ð induction, the positive cells stained by Oil Red O were 22% and 90.33%, respectively. After a 5ª²hour ¦Âª²mercaptoethanol induction of MSCs, immunohistochemistry showed that the positive rates of NSE and NFª²200 were 33% and 37.5%, and GFAP was negative. CONCLUSION: MSCs can differentiate into osteroblast, adipocyte and neurª²like cells. ......