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To further study the proliferation and multi-differentiation potentials of adipose-derived stem cells (ADSCs), the cells were isolated with the improved methods and their growth curves were achieved with cck-8. Surface protein expression was analyzed by flow cytometry to characterize the cell phenotype by PT-PCR. The multi-lineage potential of ADSCs was testified by differentiating cells with adipogenic, chondrogenic, osteogenic and myogenic inducers. The results show that about 5 × 10 5 stem cells can be obtained from 400-600 mg adipose tissue. And ADSCs can be continuously cultured in vitro for up to 1 month without passage and they have several logarithmic growth phases during the culture period. The relative surface marker expressions of all proliferated cells are positive. Meanwhile, stem cell related transcription factors, Nanog, Oct-4, Sox-2 and Rex-1, are strongly positively expressed in ADSCs. After induced differentiation toward adipogenic, osteogenic, chondrogenic and myogenic cells, the differentiated ADSCs can respectively express intracellular lipid, alkaline and calcified nodules, mucopolysaccharide-rich extracellular matrix, and a little Connexin-43, which illustrates the ability of multi-lineage differentiation of ADSCs. In order to obtain more stem cells, ADSCs are subcultured every 14 d instead of normal 5 d according to growth curve. ADSCs still keep strong proliferation ability, maintain their phenotypes, and have stronger multi-differentiation potential.

Type

Journal article

Journal

Dalian Ligong Daxue Xuebao/Journal of Dalian University of Technology

Publication Date

01/09/2010

Volume

50

Pages

649 - 655