In vitro culture of human bone marrow-derived mesenchymal stem cells and induction into osteoblasts

AIM: To build a method to culture bone marrow derived 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 calcium cobalt staining method, type Ⅰ 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 Ⅰand 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.