摘要
目的 :探讨端粒酶催化亚基 (hTERT)介导的永生化外胚间充质干细胞的生长、增殖特性及转化特征。方法 :测定永生化外胚间充质干细胞的生长曲线、群体倍增时间 ;流式细胞术检测细胞增殖周期 ;绘制细胞累计生长曲线 ;检测细胞的平板克隆形成率、软琼脂克隆形成情况 ;胰蛋白酶 -Giemsa染色法分析染色体核型 ;接种于裸鼠皮下检测细胞的致瘤性。结果 :永生化外胚间充质干细胞越过衰老期 ,增殖较活跃 ,S期细胞比例明显增加 ;该细胞不能在软琼脂内形成细胞克隆 ;第 2 5代细胞染色体形态结构正常 ,为二倍体核型 ;在裸鼠皮下不能形成肿瘤。结论 :永生化外胚间充质干细胞增殖旺盛 ,核型正常 ,无悬浮生长能力 ,在裸鼠体内不具成瘤性 ,是正常细胞而非转化细胞 ,可以作为组织工程的种子细胞。
AIM:To investigate the growth, proliferation and transforming characters of immortalized ectomesenchymal stem cells tranfected with hTERT.METHODS:The proliferation character was detected by growth curve,Flow Cytometer and flat clonogenic assay.Transforming character was detected by clonogenic soft agar assay and tumorigenic assay in nude mice.Also chromosome karyotype was analyzed.RESULTS:Immortalized ectomesenchymal stem cells proliferated actively and extended the life span beyond senescence.The cells showed normal karyotype,unable to grow in soft agar and to be nontumorigenic in nude mice.CONCLUSION:Immortalized ectomesenchymal stem cells inducted by hTERT grew actively.They were normal cells without transformed phenotype and highly desirable usage for tissue engineering.
出处
《牙体牙髓牙周病学杂志》
CAS
2004年第8期419-422,共4页
Chinese Journal of Conservative Dentistry
基金
第四军医大学创新工程重点课题 (CX0 2F0 0 2 )
关键词
外胚间充质干细胞
永生化
增殖
转化
ectomesenchymal stem cell
immortalization
proliferation
transforming
