| Objective: To investigate the effect of BMP-2 on differentiation and proliferation of cultured adipose derived adult stem cells(ADSCs)in vitro, and observe whether there is a dose-effect.Methods: Adopting three-weeks old male Sprague-Dawley rat to put to death by the methed of dislocation. Isolating adipose tissue from rat inguinal fat pads. Then digesed with collagenase-I and placed in Dulbeco Modified Eagle Medium (DMEM) with fetal bovine serum in primary cultivation. I observed the growth and appearance of the cells by a inversion microscope. When the cells are becoming stabilizing,I separated them into 8 groups (According to BMP-2 concentration, no BMP-2 group, 100ng/ml BMP-2 group,200ng/ml BMP-2 group, 400ng/ml BMP-2 group, 800ng/ml BMP-2 group, 1000ng/ml BMP-2 group, 1200ng/ml BMP-2group,1600ng/ml BMP-2 group were named respectively) and incubated in induced medium of different density of BMP-2 for 4 weeks to induce osteogenesis. Osteogenic differentiation was testd by alkaline phosphatase (ALP) staining and and type I immunohistochemistry stain. The effect of BMP-2 on differentiation and proliferation of the stem cells was compared by determining MTT activity.Results : The cells isolated from rat adipose tissue could rapidly expand. ALP staining and type I collagen immunohistochemistry staining showed positive. By contrast to OD mean of control group: 400 ng/ml BMP-2 g roup,800 ng/ml BMP-2 group , 1000 ng/ml BMP-2 group, 1600ng/ml BMP-2 had significant difference(P<0.05); 100ng/ml BMP-2 group and 200ng/ml BMP-2 group have no significant diference (P>0.05). When BMP-2 concentration varying from 100 ng/ml to 400ng/ml ,OD mean of every group increased gradually, while from 800ng/ml to 1600ng/ml, it decreased step by step.Conclusions: Adipose-derived stem cells (ADSCs) can be isolated from rat adipose tissue after osteogenesis induction. 400-800 ng/ml BMP-2 could promote differentiation and proliferation of adipose-derived stem cells significantly.
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