Human subcutaneous adipose-derived stem cells：osteoblastic/adipogenic differentiation and identification / 中国组织工程研究

ABSTRACT

BACKGROUND:Adipose-derived stem cels are a kind of mesenchyam stem cels with multipotent differentiation capacity, which have more advantages than bone marrow mesenchymal stem cels in tissue engineering research. OBJECTIVE: To establish a method to isolate and purify adipose-derived stem cels from human subcutaneous adipose tissues folowed byin vitro amplification and osteoblastic/adipogenic differentiation. METHODS: Adipose-derived stem cels were isolated from human subcutaneous adipose tissue and cultured by density gradient centrifugation and adherent culture. Cel morphology and growth features were observed under inverted microscope. Adipose-derived stem cels at passages 2 and 5 were selected for viability measurement using cel counting kit-8 method, and then cel growth curves were drawn. The immunophenotype identification was analyzed by flow cytometry. Passage 5 cels underwent osteoblastic/adipogenic induction to confirm the multi-differentiation potential. RESULTS AND CONCLUSION: (1) Using density gradient centrifugation and adherent culture method, high-purity human adipose-derived stem cels can be successfuly isolated from human adipose tissues. (2) The growth process of human adipose-derived stem cels includes stagnant phase, logarithmic phase and plateau phase, which meets the growth rhythm of normal cels. Moreover, the population doubling time is shorter. (3). Human adipose-derived stem cels are positive for stem cel-related antigens, with low immunogenicity and the multi-differentiation potential. (4) Labeling human adipose-derived stem cels with DAPI is a simple efficient labeled method, and the labeling rate is high but the cytotoxicity is low