胶原凝胶固定培养大鼠肝细胞的功能与形态观察  

Collagen-gel immobilization culture of rat hepatocytes.

作　　者：王英杰[1] 刘鸿凌[1] 文红伟[1] 刘俊[1] 郭海涛[1] 

机构地区：[1]第三军医大学西南医院,重庆400038

出　　处：《肝脏》2003年第1期15-17,共3页Chinese Hepatology

基　　金：国家自然科学基金资助项目 (39970 2 1 4 ;30 0 2 70 0 1 );全国优秀博士基金资助项目 (1 99947)

摘　　要：目的　探索混合胶原凝胶培养肝细胞的方法 ,观察培养鼠肝细胞的功能与形态特征。方法　两步法分离大鼠肝细胞 ,与Ⅰ型鼠尾胶原溶液混合接种于培养瓶 ,待胶原液形成凝胶后 ,加培养液常规培养 ,观察培养鼠肝细胞的形态学特征和尿素合成及酶漏出量。结果　成功将大鼠肝细胞混合固定于胶原中 ,形成凝胶状进行培养。培养期间 ,始终能检测出鼠肝细胞合成分泌的尿素 ,而肝细胞乳酸脱氢酶漏出量较少 ,倒置相差显微镜下观察到典型的形态特征。结论　混合胶原凝胶培养方法能为肝细胞提供更接近体内的培养环境 ,可能适用于生物人工肝研究。Objective To study the method of collagen gel immobilization culture of rat hepatocytes and to observe the function and morphological characteristics of the cultured hepatocytes. Methods Rats hepatocytes were isolated by two step perfusion and were seeded onto dishes mixed up with rat tail collagen I solution. After the collagen turned into gel form, the hepatocytes were cultured with routine culturing method. The urea(BUN) and Lactate dehydrogenase(LDH) content of the culture media and the cellular morphologies of hepatocytes in the collagen gel were observed with phase contrast photomicrographs. Results Rats hepatocytes were cultured in the collagen gel success fully. During the cultural process, the high secretion of BUN and the low leakage of LDH of the rat's hepatocytes could all be estimated. The typical cellular morphological characteristics of cultured hepatocytes could also be observed.Conclusions The collagen gel immobilization culture can provide an environment that is more closely resembing that in vivo, which maybe more eligible for use in the study of bioartificial liver.

关 键 词：肝细胞 大鼠 培养 胶原凝胶 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]