Cultivation And Identification Of Human Bone Marrow Stem Cell

Objective: To create the model of human bone marrow stromal cell(HMSCs) separation and culture in vitro. Methods: Adult bone marrow about 3~5ml were drawed by puncture,then cultured in DMEM containing Volumes of 15% Fetal bovine serum. After they grew on a great mess of surface, the adhesive cells treatment with different concentrations of dexamethasone ,cell proliferation and cell differentation were measured.And the cells were preserved to passage cultured.HBMCs were cultured in a conditional medium in subculture including dexamethasone, Vitamin C,and |3-glycerophosphate. Prolifera- tions and differentiations of all the cultured cells were observed and identificated. Results: Dexamethasone inhibited cell proliferation.With the increase of concentration of dexamethasone the effect was enhanced. At the same time dexamethasone promoted the activity of ALP.This effect was enhanced with the increase of concentration of dexamethasone. The adulthuman bone marrow stromal cells cultured in this experiment demonstrated a biological and morphologic characteristics of osteoblasts. Alkaline phosphatase(ALP) activity of HBMCs at passage 2 were 68%. The mineral nodules were observed by Von Kossa's stain, conclusion: Dexamethasone inhabited the proliferation of bone marrow stromal cells,while induced them to differentiate into osteoblasts. The appropriate concentration of dexamethasone was 10-8 mol/L.The cells cultured use the method this experiment have characteristic of osteoblasts.This method can be used as a roution method in bone tissue engineering.