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论文类型：期刊论文

发表时间：2008-08-01

发表刊物：CELL BIOCHEMISTRY AND FUNCTION

收录刊物：SCIE、PubMed、Scopus

卷号：26

期号：6

页面范围：664-675

ISSN号：0263-6484

关键字：adipose tissue; mesenchymal stem cells; proliferation; doubling time; multipoential

摘要：To further study the proliferation and multi-differentiation potentials of adipose-derived stem cells (ADSCs), the cells were isolated with improved methods and their growth curves were achieved with cck-8. Surface protein expression was analyzed by flow cytometry to characterize the cell phenotype. The multi-lineage potential of ADSCs was testified by (differentiating cells will) adipogenic. chondrogenic. osteogenic, and myogenic inducers. The results showed that about 5 x 10(5) stem cells could be obtained from 400 to 600 mg adipose tissue. The ADSCs can be continuously cultured in vitro for up to 1 month without passage and they have several logarithmic growth phases during file culture period. Also. the flow cytometry analysis showed that ADSCs expressed high levels of stem cell-related antigens (CD13. CD29, CD44. CD105. and CD166). while did not express hematopoiesis-related antigens CD34 and CD45. and human leukocyte antioen HLA-DR was also negative. Moreover, stem cell-related transcription factors. Nanog. Oct-4. Sox-2. and Rex-1 were positively, expressed in ADSCs. The expression of alkaline phosphatase (ALP) was detected in the early osteogenic induction and the calcified nodules were observed by von Kossa staining. Intracellular lipid droplets could be observed by Oil Red staining. Differentiated cardiomyocytes were observed by connexin43 fluorescent staining. In order to obtain more stem cells. we can subculture ADSCs every 14 days instead of the normal 5 days. ADSCs still keep strong proliferation ability, maintain their phenotypes, and have stronger multi-differentiation potential after 25 passages. Copyright (c) 2008 John Wiley & Sons. Ltd.