【摘要】 目的　探讨电穿孔法将ptsA5 8H质粒体外转染人胚肌腱细胞 (HETC)的条件 ,转化人胚肌腱细胞 (THETC)体外培养的生长特性。方法　体外分离培养HETC ,电穿孔法导入 ptsA5 8H质粒 ,并经潮霉素B筛选 ,获得阳性克隆 ,扩增后建立THETC系。体外传代培养、冻存、复苏THETC ,并分别绘制HETC和THETC在不同培养条件的生长曲线。结果　通过潮霉素B 0～ 10 0 0mg/L终浓度梯度的培养 ,选择 2 0 0mg/L作为筛选浓度。电穿孔法击穿电压选择 96 0 μF、40 0V。THETC增殖能力强 ,可长期连续传代 ,冻存、复苏不改变其生长特性 ,体外培养控制温度可改变其生长特性 ,且其生长存在血清营养依赖性。结论　将 ptsA5 8H质粒转入人胚肌腱细胞可使肌腱细胞在体外长期连续传代 ,为组织工程化肌腱的研究提供标准细胞。更多还原

【Abstract】 Objective To explore the transfection conditions of human embryonic tendon cells (HETC) by ptsA58H plasmid, and to investigate the growth characteristics of transformed human embryonic tendon cells (THETC). Methods The human embryonic tendon cells were isolated and cultured in vitro . By using the technique of electroporation, a plasmid ptsA58H was transformed into HETC, and the positive clones were formed following strict selection by hygromycin B. THETC were subcultured and their growth curves were drawn in different culture conditions.Results Through concentration grading culture of hygromycin B(0～1 000 mg/L), the optimal screening concentration was 200 mg/L, and the optimal voltage was 960 uF and 400 V. The proliferation capability of THETC was strong and THETC could be subcultured continually and permanently. The growth properties of THETC were not changed when cells were frozen storage. The growth characteristics of THETC could be controlled when the culture temperature was changed. Moreover, the growth of THETC was serum-dependent.Conclusion HETC can be cultured continuall and permanently after transformed by ptsA58H plasmid, it is the ideal standard cell line for the research of tissue engineered tendon.更多还原