上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 技术与方法 • 上一篇    

正常大鼠肝脏祖细胞的分离和培养

臧金锋1,2，袁 寅2，高军业2，赵 燕3，钱海鑫1   

  1. 1.苏州大学附属第一医院普外科， 苏州 215006； 2.泰州市人民医院肝胆外科， 泰州 225300； 3.泰州市人民医院病理科， 泰州 225300
  • 出版日期:2014-05-28 发布日期:2014-05-30
  • 作者简介:臧金锋（1979—）， 男， 副主任医师， 博士生; 电子信箱: xkfy1979@126.com。

Isolation and in vitro culture of hepatic progenitor cells of normal rat liver

ZANG Jin-feng1,2, YUAN Yin2, GAO Jun-ye2, ZHAO Yan3, QIAN Han-xin1   

  1. 1.Department of General Surgery, the First Affiliated Hospital of Soochow University, Suzhou 215006, China; 2.Department Hepatobiliary Surgery, People's Hospital of Taizhou, Taizhou 225300, China; 3.Department of Pathology, People's Hospital of Taizhou, Taizhou 225300, China
  • Online:2014-05-28 Published:2014-05-30

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摘要：

目的 探讨正常大鼠肝脏祖细胞的分离和体外培养方法。方法 选择正常Wistar大鼠，以CD90.1为标志物，通过免疫磁珠法分离正常肝脏内的祖细胞。对祖细胞进行体外培养，并诱导分化。结果 CD90.1阳性细胞占肝脏非实质细胞的（0.32±0.03）%，流式细胞仪分析显示CD90.1阳性细胞占分离细胞的（98.26±1.37）%。刚分离的CD90.1阳性细胞中，CK-19表达呈强阳性，HNF-4a表达呈阴性；培养8周后，CK-19表达呈阴性，HNF-4a表达呈阳性。结论 正常大鼠肝脏内存在祖细胞，可将其诱导分化为肝细胞。

关键词: 大鼠, 肝脏祖细胞, 细胞分选, 细胞培养

Abstract:

Objective To explore the techniques of isolation and in vitro culture of hepatic progenitor cells of normal rat liver. Methods According to the cell surface marker CD90.1, hepatic progenitor cells of normal Wistar rats were isolated by immunomagnetic cell sorting, cultured in vitro, and induced to differentiate. Results The percentage of CD90.1 positive cells in non-parenchymal cells was (0.32±0.03) %. The flow cytometric cell sorting showed that (98.26±1.37) % of isolated cells were CD90.1 positive cells. Fresh isolated CD90.1 positive cells were strong positive for CK-19 and negative for HNF-4a. After 8 weeks of culture, these cells were negative for CK-19 and positive for HNF-4a. Conclusion Hepatic progenitor cells exist in normal rat livers and can be differentiated toward hepatocytes.

Key words: rat, hepatic progenitor cell, cell sorting, cell culture