【摘要】 目的研究人脂肪间充质干细胞(ADSCs)在体外传代过程中生物学特性及遗传特性的变化,为脂肪间充质干细胞的基础研究及临床应用安全性提供理论依据和新的细胞来源。方法利用无血清培养基培养ADSCs,对1、3、5、7、10、14和15代体外培养细胞进行了形态观察,流式细胞术分析细胞周期及免疫表型,G显带法分析染色体核型,实时荧光定量分析相关基因表达量,ELISA定量分析细胞因子的表达量。结果 ADSCs传代至第7代后,细胞增殖速度开始减慢,10代后细胞增殖速度显著减慢。细胞形态为长梭形并保持稳定。处于G2期和S期细胞百分比基本保持稳定。染色体细胞核型没有明显易位和缺失等变化,相关基因表达量略有差异,培养基内分泌的细胞因子保持稳定。结论利用无血清培养基培养的脂肪间充质干细胞在体外培养5代以内基本安全。更多还原

【Abstract】 Objective To investigate the biological characteristics and genetic features alteration of human adipose derived mesenchymal stem cells( ADSCs) cultured in vitro and to provide a new source of cells for the research on the safety of its clinical trials. Methods Observe the shape of the 1 st,3 rd,5 th,7 th,10 th,14 th and15 th generation of the ADSCs cultured in vitro using the serum free culture medium. Analyze their cell cycle and cell surface markers by flow cytometry,karyotype was analyzed by the chromosome G-banding technology. Expression level of relevant gene and cytokine was Measured respectively by the Real-time fluorescence quantitative technique and ELISA. Results The growth of ADSCs began to slow down after the 7 th generation,and after the10 th generation,it slowed down significantly. The shape of cell remained fusiform,no significant change. Cellcycle analysis revealed that the percentage of G2 phase and S phase cells remained stable. There was no obvious missing,translocation or dislocation in terms of karyotype. Measured gene expression level had a little difference change,and the cytokine secreted in culture medium remained stable. Conclusions The adipose derived mesenchymal stem cells cultured in vitro using the serum free culture medium are safe before the 5th generation.更多还原