摘要
目的研究小鼠胚胎干细胞（embryonicstemcells，ESC）诱导分化的血管内皮细胞永生化后的致瘤现象。方法体外培养系中，将诱导的小鼠ESC的拟胚体分化为圆形细胞。通过脂质体将人端粒酶催化亚基（human telomerase reverse transeriptase，hTERT）基因转染诱导分化中的圆形细胞，应用RT—PCR及免疫组织化学等方法分析、观察诱导分化的血管样结构的组成细胞和被hTERT基因转染的圆形细胞的形态和生物学特性。并将基因转染的细胞植入裸鼠皮下，观察其致瘤性。结果携带hTERT基因、从ESC分化而来的圆形细胞TEC3，在体外可大量增殖，培养的前2d（48h），细胞数就增加了1倍多，至72h细胞数增加了12倍，持续传代，端粒酶活性高表达，95％细胞表达Flk-1、CD34和vWF，并有管道化生长特性。将细胞植入裸鼠皮下后1周有肿瘤形成。结论转染hTERT基因可使小鼠ESC分化细胞持续增殖，植入裸鼠体内具有致瘤性。
Objective To discuss the tumorigenicity of immortalized endothelial cells differentiated from embryonic stem cells. Methods The embryoid bodies （EB） formed in vitro from embryonic stem cells, were induced to differentiate into many ＂round cells＂ （ the precursor of endothelial cells）. These ＂round cells＂ later formed the vascular tube-like structures. To immortalize these cells, human telomerase reverse transcriptase （hTERT） cDNA was transfected into ＂round cells＂ by lipofectine, RT-PCR and immunocytochemistry were used to evaluate the immortalized cells. And the tumorigenicity of these cells were evaluated by being injected into nude mice subcutaneously. Results 95% of these transfected cells expressed Flk-1, CD34 and vWF, and could proliferate in large quantity in vitro （ cell number was doubled in 2 days, and increased 12 times in 3 days） , and were able to form tubular structures. Conclusions These results suggest that hTERT cDNA transfection can immortalize induced endothelial cells and tumorigenicity is found after immortalizated cells are injected into nude mice subcutaneously.
出处
《中华整形外科杂志》
CAS
CSCD
北大核心
2012年第1期33-39,共7页
Chinese Journal of Plastic Surgery
基金
基金项目：国家自然科学基金面上项目（81071480）
关键词
胚胎干细胞
内皮细胞
人端粒酶催化亚基基因
致癌性试验
Embryonic stem ceils
Endothelial cells
Human telomerase reverse transcriptase
Carcinogenicity tests
