转化人胚肌腱细胞的体外培养及生物学特性研究  

CULTURE OF THE TRANSFORMED HUMAN EMBRYONIC TENDON CELLS AND ITS BIOLOGICAL CHARACTERISTICS IN VITRO

作　　者：杨志明[1] 解慧琪[1] 项舟[1] 李胜富[1] 魏启欧[1] 

机构地区：[1]华西医科大学附属第一医院骨科

出　　处：《中国修复重建外科杂志》1999年第2期99-104,共6页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金;美国纽约中华医学基金

摘　　要：目的研究经ｐｔｓＡ５８Ｈ质粒转化的人胚肌腱细胞的生物学特性，探讨细胞生长特性与体外培养条件改变的关系。方法取人胚肌腱细胞和经ｐｔｓＡ５８Ｈ质粒转化的人胚肌腱细胞，进行体外培养。对细胞进行形态学、超微结构及一般特征的观察。在不同培养条件下进行生长曲线绘制，平板克隆实验和软琼脂培养以及免疫组织化学法胶原染色。结果在正常培养条件下，人胚肌腱细胞和转化人胚肌腱细胞的生长性状几乎无差异，且冻存复苏并不改变其生长特性。在３９．５℃及３５℃进行条件培养，转化细胞的生长特性发生改变。转化细胞能长期传代，增殖能力强，并存在接触抑制现象和血清营养依赖性，在软琼脂中不能生长，具有分泌Ⅰ型胶原的能力。结论经ｐｔｓＡ５８Ｈ质粒转化的人胚肌腱细胞可长期连续传代。通过改变培养条件可控制其生长。无肿瘤细胞的生物学特点。Objective This paper was to study the biological characteristics of the transformed human embryonic tendon cells, the relation between cell growth and culture conditions, and to compare these features with that of human embryonic tendon cells. Methods The pts A58H plasmid had successfully used to transform a tendon cell line from human embryo in our past work. The human embryonic tendon cells and the transformed human embryonic tendon cells were cultured in vitro. In different culture conditions, the growth curve were drawn respectively. Population dependence and proliferation capability of the cells were investigated through plate cloning test and soft agar culture. The collagen secreted by cells was identified by immunohistochemical method. Results In routine culture condition, the growth properties of the human embryonic tendon cell and transformed cells were almost identical. The growth properties of the transformed cells were not changed when the cells were frozen storage. There were changes of growth characteristics of the transformed cells when the culture temperature was changed. The transformed cells could subcultured continually and permanently. The proliferation capability of the transformed cells were stronger than that of the human embryonic tendon cells. Moreover, the growth of the transformed cells was serum dependent, and the phenomenon of contact inhibition was observed. The transformed cells were not able to grow on soft agar culture. They had the capacity of secreting collagen type Ⅰ. Conclusion The transformed human embryonic tendon cells could be subcultured continually and permanently, and their growth could be controlled by changing their culture conditions and they had no malignant tendency in biological characteristics. They could be taken as an ideal experimental material for tendon engineering.

关 键 词：人胚肌腱细胞 转化细胞 体外培养 生物学特性 

分 类 号：R329－33[医药卫生—人体解剖和组织胚胎学]