In vitro culture of human bone marrowª²derived mesenchymal stem cells and induction into osteoblasts

    TENG Yong, HU Yunª²Yu, AN Suª²Jie, DAI Xianª²Wen, ZAO Li, BAI Jianª²Ping, LI Xuª²Sheng, L¨¹ Rong

    1Department of Orthopaedics, Xijing Hospital, 2College of Postgraduate, Fourth Military Medical University, Xi¬ðan 710033, China

    ¡¾Abstract¡¿ AIM: To build a method to culture bone marrowª²derived mesenchymal stem cells (MSCs) in vitro and to investigate the feasibility of inducing MSCs into osteoblasts. METHODS: Human MSCs were isolated from adult bone marrow and purified by percoll density gradient centrifugation and cultured in vitro. The MSCs attachment formed after 7-10 d and the MSCs of the passage 3 were chosen to induce into osteoblasts. Fifteen days later, the alkaline phosphatase assay was conducted using modified calciumª²cobalt staining method, type ¢ñ collagen and osteocalcin assay was conducted using immunohistochemistry and calcium node assay was done using alizarin red staining. RESULTS: Homogeneous MSCs were obtained by percoll density gradient centrifugation. The induced MSCs had typical appearance of osteoblasts. The rate of ALP expression was 81%, the expressions of collagen type ¢ñand osteocalcin were positive, and calcium nodes were seen. CONCLUSION: The MSCs can be separated from adult bone marrow and cultured in vitro, which can be induced into osteoblasts. ......