Study on adipose tissue-derived stem cells of rat differentiating into the osteoblast in vitro

AIM:To explore the capability of adipose tissue-derived steme cells differentiating into osteoblast in vitro, to evaluate the feasibility of ADSCs as seeding cells in bone tissue engineering.METHODS:The adipose tissue were isolated from the inguinal fat pads of the SD rats . Primary ADSCs were obtained by the methods of collagenaseⅠdigestion. Subsequently, the ADSCs were cultured and subcultured. Cell morphology and proliferation characteristics were observed under inverted phase contrast microscope every day. The third passage of ADSCs were induced into osteoblasts by the osteogenic inducing fluid. The osteogenic phenotype were examined by means of alkaline phosphatase(ALP),Von Kossa stainings and immunocytochemistry. RESULTS:ADSCs were successfully obtained and cultured from rat adipose tissue. They appeared fibroblast-like and could proliferate rapidly in vitro. After induced with dexamethasone, ascorbic acid, beta-sodium glycerophosphate and 1,25-(OH)_2Vit D_3, ADSCs epressed the characteristic of osteoblasts: increasing of alkaline phosphatase(ALP) actiuity significantly, appearance of calcium node showed under Kossa staining and positive OC、collagenⅠthrough immunocytochemistry. CONCLUSION:Rat ADSCs could be obtained easily. They have a strong capability of proliferation and osteogenic properties. This study showed that the rat ADSCs could be used as seed cells in the bone tissue engineering.