Isolation,culture and osteogenic differentiation of rat adipose-derived stem cells

Objective: To study the biological characteristics and osteogenic potency of adipose-derived stem cells(ASCs).Methods: Cells were isolated from the bilateral inguinal fat pads by collagenase I digestion and cultured.Cell morphology was observed under the phase contrast microscope.The cell proliferation was tested by MTT assay.The surface markers of the cells were analyzed by flow cytometry.Passage 3 cells were cultured in osteogenic and adipogenic inducing medium for 7,28 and 21 days respectively,and verified with ALP staining,Alizarin red staining and Oil Red O staining respectively.Results: The cells presented fibroblast-like or polygon shape.The growth curve was like "S" shape reflecting strong proliferative ability.ASCs expressed CD29 and CD90,but not CD34 and CD45.The ALP staining,Alizarin red staining and Oil Red O staining showed positive reaction.Conclusion: ASCs can be cultured from SD rat fat tissue and can be induced to osteogenic differentiation.