摘要
目的分离及培养高纯度的原代大鼠肝细胞。方法用D-Hanks液及胶原酶消化液分两步原位灌注大鼠肝脏,分离细胞经3次低速离心(50×g,5 min)后获得纯化的肝细胞。肝细胞存活率检测用台盼蓝拒染法,鉴定用PAS染色法。结果每只体质量约200 g的大鼠能成功分离出(1.0～1.5)×108个肝细胞,成活率在90%以上,细胞培养24 h,有部分细胞贴壁,48 h有大量细胞贴壁。以PAS染色对肝细胞进行鉴定,肝细胞为粉红色。结论肝细胞分离培养成功,对分离方法的改进措施可行。
Objective To establish a method of separation and culture of high purity primary rat hepatocyte. Methods The perfusion solutions used in the first and the second step were D-Hankg solution and the digestive solution contained eollagenase Ⅳ, respectively. Purified hepatocytes were separated from the dissociative cells by low-speed centrifugation （50 x g, 5 rain） 3 times. The survival rate was measured by typan blue exclusion and the identification of hepatocytes was measured by PAS cytochemistry. Results （ 1.0 - 1.5） x 10^8 hepatocytes were prepared from about a 200 g rat on average. The survival rate was higher than 90%. Part of the cells adhered in 24 h and much of them adhered in 48 h. The cells by PAS cytochemistry were identified. The cells were pink. Conclusion The rat hepatocytes are success- fully isolated and cultured.
出处
《胃肠病学和肝病学杂志》
CAS
2011年第6期568-570,共3页
Chinese Journal of Gastroenterology and Hepatology
关键词
肝细胞
分离
培养
Hepatocyte
Separation
Culture
