标题：小鼠原代肝细胞的分离与培养

      作者：田守润 1，勾艳芳 1，王锦哲 1，樊芮伽 1，唐历波 2    1.海南医学院第二临床学院，海南 海口 570145；2.海南医学院基础医学与生命科学院，海南 海口 570145

      卷次： 2022年33卷18期

      【摘要】 目的 探索小鼠原代肝细胞细胞分离培养的简化方法，并对其生物学特性进行鉴定。方法 分离出生2~3 d小鼠肝组织，利用低温下机械剪碎法结合胶原酶消化，分离小鼠原代肝细胞，利用小鼠肝表面抗原CK-19进行荧光染色鉴定。结果 成功地从乳鼠体内分离获得肝原代细胞，免疫荧光鉴定属于肝细胞。结论 利用简单的机械加酶消化法也可以获得肝组织细胞，且得率较高，细胞活力较好。
      【关键词】 原代肝细胞；培养鉴定；胶原酶消化法；荧光染色；鉴定；细胞培养
      【中图分类号】 R-332 【文献标识码】 A 【文章编号】 1003—6350（2022）18—2317—04

Isolation and culture of primary hepatocytes in mice.
TIAN Shou-run 1, GOU Yan-fang 1, WANG Jin-zhe 1, FANRui-jia 1, TANG Li-bo 2. 1. The Second Clinical College of Hainan Medical University, Haikou 570145, Hainan, CHINA;2. College of Basic Medicine and Life Sciences, Hainan Medical University, Haikou 570145, Hainan, CHINA
【Abstract】 Objective To explore a simplified method for the separation and culture of primary hepatocytes inmice. Methods The liver tissues of 2-3-day-old mice were isolated, and the primary hepatocytes of mice were isolatedby mechanical shredding at low temperature combined with collagenase digestion. The fluorescent staining identificationwas carried out by using mouse liver surface antigen CK-19. Results The primary liver cells were successfully iso-lated from suckling mice, and they were identified as liver cells by immunofluorescence. Conclusion The mechani-cal shredding and enzymatic digestion method can also be used to obtain liver tissue cells, with high yield and goodcell viability.
      【Key words】 Primary hepatocytes; In vitro isolation and culture; Collagenase digestion; Fluorescence staining;Identification; Cell culture