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论文类型：期刊论文

发表时间：2012-12-01

发表刊物：APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY

收录刊物：Scopus、PubMed、SCIE、EI

卷号：168

期号：8

页面范围：2230-2244

ISSN号：0273-2289

关键字：Human adipose-derived stem cells; Growthcurves; Cellcycle; CyclinA; CyclinD1

摘要：Abundant and less passaged cells are highly expected in clinical application since repeated subculture reduces stem cell characteristics. Long time culture of stem cells without passage is therefore needed. The growth and cell viability of human adipose-derived stem cells (hADSCs) were investigated by live/dead staining, cck-8 kits, and hemocytometer every day in 30 days of culture. The stem cell characteristics of hADSCs at the beginning and the end of culture were detected by flow cytometry and histochemical staining. hADSCs can be cultured up to the 30th day in one passage while maintaining high level cell viability and their stem cell characteristics. In addition, the cells displayed two plateau phases and three logarithmic phases during 1 month of culture. Increasing expression of cyclin A at protein level resulted in an increase in the percentage of hADSCs in the S and G(2)/M phases, while decreasing protein level of cyclin D1 induced a decline in the proportion of hADSCs in the G(0)/G(1) phase, regulating cells to move into rapid proliferation. This study demonstrates that a great quantity of hADSCs can be obtained in vitro by prolonging the culture time of each passage. And cyclin A and cyclin D1 affect the distribution of cell cycle and regulate the growth of hADSCs.