Osteoblast-differentiation of rabbit adipose-derived stem cells by in vitro transfection of bone morphogenetic protein-7

Abstract

Objective To explore the osteogenic potential of adipose-derived stem cells (ADSCs)when exposed to the cDNA of human bone morphogenetic protein-7 (hBMP-7) during osteoblast-differentiation in vitro. Methods The ADSCs were primarily cultured from rabbit fat tissue. CD44, CD49d and CD106 were detected and identified by immunohistochemistry. ADSCs were inducted with the method of stable transfection of pcDNA3. 1-GFP-hBMP-7 by Liprofectamine2000. The green fluorescent protein (GFP) expression was observed to access transfection efficiency of the transfected cells. RT-PCR was used to detect the expression of hBMP-7 gene in the induced ADSCs. The transform of ADSCs was assessed by detecting the ALP and expressions of collagen Ⅰ and osteocalcin. Results ADSCs were successfully isolated from rabbit adipose tissue. The isolated ADSCs expressed CD44 and CD49d but CD106 was absent. The pcDNA3. 1-GFP-hBMP-7 transfected ADSCs expressed hBMP-7. The intracellular ALP spectrophotometry in the hBMP-7 group was higher than in the control group after transfection. The expressions of collagen Ⅰ and osteocalcin in the hBMP-7 group were higher than in the control group. Conclusion ADSCs isolated from rabbit adipose tissue may be an excellent seeding cell for tissue engineering because they can differentiate into osteoblasts in vitro by hBMP-7 gene transfection. Key words: Adult stem cells; Bone morphogenetic proteins; Transfection; Cell differentiation