摘要
目的研究体外贴壁培养肌腱细胞的功能老化。方法取材成年鸡的趾深屈肌腱,经胶原酶消化法获取肌腱细胞,在体外进行传代培养,收集不同培养时期的细胞,进行形态学、细胞增殖、组织学、逆转录聚合酶连反应(RT-PCR)及Westernblot技术分析,观察不同体外培养时间肌腱细胞表型的变化。结果组织学、RT-PCR及Westernblot等均证实随着体外培养时间的延长,肌腱细胞传至第5代后,细胞形态发生明显改变,细胞增殖能力显著减弱,胶原分泌明显减少。结论体外贴壁培养的肌腱细胞传至第5代后,细胞表型发生显著老化的改变。
Objective To explore a feasible method to fabricate tissue-engineered tendon in im munocompetent animals by tissue engi-neering approach,study on characte ristics of cultured tenocytes in vit ro.Methods Chickens were used in this study.Ten dons obtained from the third digital deep flexor tendon were digested by typeⅡcollagenase .The tenocytes were harvested and purified every hour,cult ured in vitro.A series of studies were conducted.We detected collagen in tenocy tes by Masson' trichrome,the expression of mRNA by RT-PCR,contents of type I collagen in cell cu lture medium by Western blot to evalu ate tenocytes viability.Results Prolonged the culture in vitro,phenotype of tenocytes altered and t heir ability to secrete collage decreased,especially distinct after th e fifth passage.Conclusions Tenocytes cultured in vitro underwent senesce nt procedure with changing in their p henotype,obviously after the fifth passage.
出处
《中国临床康复》
CSCD
2002年第22期3376-3377,I003,共3页
Chinese Journal of Clinical Rehabilitation
基金
上海市科学技术委员会基金资助项目(974119008)
