EXPERIMENTAL STUDY OF CULTIVATION,IDENTIFICATION AND INDUCED DIFFERENTIATION OF BONE MARROW STROMAL STEM CELLS INTO OSTEOBLASTS IN RATS

Objective To set up a protocol of culturing bone marrow stromal stem cells(BMSCs) of rats in vitro,and to study the condition for induced differentiation to osteoblasts. Methods BMSCs were isolated from adult rats for anchoring culture.The passage cells were induced to differentiate into osteoblasts in a differentiation induction medium.The phenotype was determined by morphological method,and levels of alkaline phosphatase(ALP) and osteocalcin(OCN),and ability to form mineralized nodules were examined. Results Colonies of BMSCs were formed in the primary culture and contacted to each other at day 14.In the passage culture,the cells became bigger than those in the primary culture and a new generation was produced in 5—7 days.Under induction,levels of ALP were elevated and the morphological changes of osteoblast,phases of proliferation,aggregation,nodulation and mineralization were observed.During the mineralization,expression of OCN was detected. ConclusionBMSCs can be easily isolated and cultured in vitro,with strong proliferation ability and osteogenic capacity.BMSCs can be used as seed cells for tissue engineering of bone.