ptsA58H质粒转化人胚腱细胞的生物学特性研究

Experimental study on the biological characteristics of ptsA58H plasmid transformed human embryonic tenocytes

目的 研究经ptsA5 8H质粒转染的转化人胚腱细胞的生物学特性及致瘤性。方法 体外培养转化人胚腱细胞 ,对细胞进行形态学、超微结构及一般特征的观察。绘制细胞生长曲线 ,平板克隆实验检测克隆形成率 ,软琼脂培养 ,植入裸鼠体内观察有无致瘤性 ,Brdu标记细胞 ,检测细胞植入体内的转归及细胞的分泌功能。结果 转化人胚腱细胞能长期传代 ,增殖能力强 ,在软琼脂中不生长 ,接种裸鼠不致瘤 ,且在裸鼠体内细胞存活 ,增殖能力旺盛 ,具有正常分泌Ⅰ型胶原的功能。结论 经ptsA5 8H质粒转化的人胚腱细胞可长期连续传代 ,增殖能力强 ,无致瘤性 ,可作为组织工程化肌腱的标准化细胞。

Objective To study the biological characteristics and oncogenicity of human embryonic tenocytes transformed by ptsA58H plasmid. Methods The transformed human embryonic tenocytes (THETC) were cultured in vitro. Their morphological characteristics and ultrastructure were observed. Clone efficiency was detected by plate cloning test. Oncogenicity was investigated by soft agar culture and transplantation to nude mice. THETC were labeled by Brdu to examine the conversion and secretion activity. Results THETC could be subcultured continuously with strong proliferation ability. They could not grow in soft agar. No oncogenicity after transplantation to nude mice was observed. The cells could survive and proliferate in nude mice and secret type Ⅰ collagen. Conclusions The human embryonic tenocytes transformed by ptsA58H plasmid can be subcultured continuously and permanently, with strong proliferation ability, and no oncogenicity. They can be used as the ideal standard cells for the tissue engineered tendon.