摘要
研究了脂肪干细胞在一个月连续培养中的生长增殖特性,及经长期连续培养后细胞的干细胞特性,同时探索了细胞周期蛋白表达与生长增殖特性之间的关系。分别采用CCK-8试剂盒与血细胞计数板来检测细胞活性与细胞数随时间的变化;用流式细胞仪检测细胞的死活情况;采用流式细胞仪和组织化学染色分析经一个月连续培养后细胞的干细胞特性;流式细胞仪和western blot分析不同时间段的细胞周期分布和细胞周期蛋白的表达。结果表明,脂肪干细胞在一个月的连续培养过程中始终保持很高的细胞活性和增殖能力。并且连续一个月培养后,仍然保持其干细胞特性。细胞周期蛋白A的增量表达,促进脂肪干细胞从S期向G2/M期过渡,使G2/M期细胞比率增加,处于分裂状态细胞含量增加,此时细胞处于快速增殖状态。因此,可以通过延长脂肪干细胞的传代培养时间来为细胞治疗提供更多高质量的移植用干细胞。并且在脂肪干细胞长期连续培养过程中,细胞周期蛋白A通过调节细胞周期分布而影响着细胞的生长增殖。
A great number of human adipose-derived stem cells （ADSCs） are required for cell therapy, it isthus necessary to expand ADSCs and investigate their proliferation characteristics in vitro in a long-term culture.The growth characteristics of ADSCs were investigated by CCK-8 kits and hemocytometer everyday during 30days culture. The stem cell characteristics of ADSCs after one month culture were detected by flow cytometry（FCM） and histochemical staining. The cell cycle distribution profiles and the expression of cyclin A wererespectively detected by FCM and western blot. The results demonstrate that ADSCs maintain high proliferationcapacity and cell viability during one month culture. In addition, these cells still maintain their characteristics ofstem cells after one month culture. Increasing expression of cyclin A at protein level promotes the transitionfrom S phases to G2 / M phases, and results in increasing the percentage of ADSCs in G2/M phases. The cellsthus proliferate rapidly. Prolonging the culture time can be adopted to obtain higher quality stem cells used forthe cell transplanting of the cell therapy. In addition, cyclin A affects the proliferation of ADSCs by regulatingthe distribution of cell cycle during long time culture.
出处
《高校化学工程学报》
EI
CAS
CSCD
北大核心
2014年第2期275-281,共7页
Journal of Chemical Engineering of Chinese Universities
基金
国家自然科学基金(31170945)
