中华整形外科杂志
中華整形外科雜誌
중화정형외과잡지
CHINESE JOURNAL OF PLASTIC SURGERY
2012年
1期
33-39
,共7页
胚胎干细胞%内皮细胞%人端粒酶催化亚基基因%致癌性试验
胚胎榦細胞%內皮細胞%人耑粒酶催化亞基基因%緻癌性試驗
배태간세포%내피세포%인단립매최화아기기인%치암성시험
Embryonic stem cells%Endothelial cells%Human telomerase reverse transcriptase%Carcinogenicity tests
目的 研究小鼠胚胎干细胞(embryonic stem cells,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 transcriptase,hTE RT)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2 d(48 h),细胞数就增加了1倍多,至72 h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.
目的 研究小鼠胚胎榦細胞(embryonic stem cells,ESC)誘導分化的血管內皮細胞永生化後的緻瘤現象.方法 體外培養繫中,將誘導的小鼠ESC的擬胚體分化為圓形細胞.通過脂質體將人耑粒酶催化亞基(human telomerase reverse transcriptase,hTE RT)基因轉染誘導分化中的圓形細胞,應用RT-PCR及免疫組織化學等方法分析、觀察誘導分化的血管樣結構的組成細胞和被hTERT基因轉染的圓形細胞的形態和生物學特性.併將基因轉染的細胞植入裸鼠皮下,觀察其緻瘤性.結果 攜帶hTERT基因、從ESC分化而來的圓形細胞TEC3,在體外可大量增殖,培養的前2 d(48 h),細胞數就增加瞭1倍多,至72 h細胞數增加瞭12倍,持續傳代,耑粒酶活性高錶達,95％細胞錶達Flk-1、CD34和vWF,併有管道化生長特性.將細胞植入裸鼠皮下後1週有腫瘤形成.結論 轉染hTERT基因可使小鼠ESC分化細胞持續增殖,植入裸鼠體內具有緻瘤性.
목적 연구소서배태간세포(embryonic stem cells,ESC)유도분화적혈관내피세포영생화후적치류현상.방법 체외배양계중,장유도적소서ESC적의배체분화위원형세포.통과지질체장인단립매최화아기(human telomerase reverse transcriptase,hTE RT)기인전염유도분화중적원형세포,응용RT-PCR급면역조직화학등방법분석、관찰유도분화적혈관양결구적조성세포화피hTERT기인전염적원형세포적형태화생물학특성.병장기인전염적세포식입라서피하,관찰기치류성.결과 휴대hTERT기인、종ESC분화이래적원형세포TEC3,재체외가대량증식,배양적전2 d(48 h),세포수취증가료1배다,지72 h세포수증가료12배,지속전대,단립매활성고표체,95％세포표체Flk-1、CD34화vWF,병유관도화생장특성.장세포식입라서피하후1주유종류형성.결론 전염hTERT기인가사소서ESC분화세포지속증식,식입라서체내구유치류성.
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.
