Cultivation and identification of bone mesenchymal stem cells of ovariectomied SD rats in vitro and characters of proliferation
AIM: To isolate,culture and identify bone marrow stromal cells (BMSCs) of ovariectomied Sprague-Dawley(SD) rats and to study their proliferation features. METHODS: Three-month-old SD rats were used in this study. Twelve female rats were randomly divided into 2 groups,6 rats were ovariectomied as experimental group Ⅰ while other 6 rats received sham-operation as group Ⅱ. After 3 months,BMSCs of the 12 rats were isolated by gradient centrifugation as primary culture and the isolated BMSCs were identified by flow cytometer and multi-differentiation cultivation. The cells were screened by inverted microscope each day and MTT assay was performed to determine the cell proliferative activity. RESULTS: The weight,estrogen and bone mineral density of the rats were statistically different between experimental group and control group. Von Kossa's staining demonstrated positive osteoblast phenotypes of mineralized nodes by osteogenic inducer and Oil Red 0 staining showed that BMSCs treated with adipogenic medium resulted in adipocyte formation. Flow cytometer showed positive expression of CD29 and CD44 and negative expression of CD45 and CD133. MTT detection displayed that the proliferation activity of the cells in experimental group was higher than that in control group every day within 7 d. CONCLUSION: The model of osteoporosis has been successfully established. The BMSCs isolated from the SD rats with osteoporosis have multipo-tential ability,and increased protiferative activity.
ovariectomyratsbone marrow stromal cellculture in vitroproliferation
NETL
NSTL
万方数据
维普
