简易胶原酶消化法提取兔原代肝细胞及培养  

Isolation and culture of rabbit primary hepatocytes using simple collagenase digestion method

作　　者：唐靓[1] 罗涛[2] 杨亚冬[2] 张文元[2] 

机构地区：[1]浙江省医学科学院保健食品研究所,杭州310013 [2]浙江省医学科学院生物工程研究所,杭州310013

出　　处：《现代医药卫生》2017年第12期1764-1765,1768,共3页Journal of Modern Medicine & Health

基　　金：浙江省科技计划公益技术研究社会发展项目(2015C33109);浙江省科研院所专项基金资助项目(2015F10014;2016F10002);浙江省医药卫生科技计划项目(2016KYB071;2017KY299)

摘　　要：目的探讨简易、经济的Ⅳ型胶原酶消化法提取分离新生兔原代肝细胞及体外培养。方法采用0.1%Ⅳ型胶原酶对新生兔肝脏组织小块进行消化以提取兔原代肝细胞,体外进行单层培养,并对提取的兔肝细胞的细胞数量、细胞活力和细胞增殖能力进行检测。结果提取的肝细胞主要呈类圆形,排列整齐,细胞透亮有光泽,立体感好,细胞界限清晰。1 g肝组织可分离1.12×10~6个肝细胞,成活率为77%。细胞增殖能力呈先升后降趋势,细胞生长曲线较符合原代细胞生长的一般规律。结论该实验方法简单、易操作,无需特殊设备,适合一般实验室开展原代肝细胞的提取、分离与培养。Objective To explore isolation and culture of primary hepatocytes from newborn rabbit by a simple and economical method of collagenase digestion. Methods Rabbit primary hepatocytes were extracted from liver tissue pieces of newborn rabbit by 0.1% collagenase type Ⅳ digestion, and were Cultured in vitro. Then the cells number, cell viability, and cell proliferation ability were detected. Results The primary hepatocytes were round-like,neat rows,bright luster,with good stereo sense, and clear cell lines. 1.12× 10^6 hepatocytes could be extracted from 1 g liver, and survival rate of the hepatocytes was 77%. Cell proliferation capacity showed a tendency of rising first and falling later, and cell growth curve corresponds to general rule of primary cells growth. Conclusion The method is simple, easy to operate, and no need special equipment, which is suitable for general laboratory to carry out extraction, isolation, and culture of primary hepatocytes.

关 键 词：原代肝细胞 Ⅳ型胶原酶 细胞提取 细胞培养 新生兔 

分 类 号：R657.3[医药卫生—外科学]