Simultaneous isolation and primary culture of rat hepatocytes, hepatic stellate cells, Kupffer's cells and hepatic sinus endothelial cells
10.3969/j.issn.1673-4254.2014.04.20
- VernacularTitle:大鼠原代肝细胞、星状细胞、枯否细胞和肝窦内皮细胞的同步分离与培养
- Author:
Yang LI
1
;
Shuangming CAI
;
Lili ZHANG
;
Xu LI
Author Information
1. 南方医科大学南方医院急诊科
- Keywords:
primary culture;
hepatocytes;
hepatic stellate cells;
Kupffer's cells;
hepatic sinus endothelial cells;
in situ perfusion;
density gradient centrifugation
- From:
Journal of Southern Medical University
2014;(4):532-537
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a method for simultaneous isolation and primary culture of rat hepatocytes, hepatic stellate cells, Kupffer's cells and hepatic sinus endothelial cells. Methods By combining in situ perfusion, in vitro perfusion, density gradient centrifugation and differential adhension, primary rat hepatocytes, hepatic stellate cells, Kupffer's cells and hepatic sinus endothelial cells were obtained. The purity of these cells were assessed with morphological observation, immunofluorescent staining and ink phagocytosis assay. Results We successfully obtained the 4 primary cells simultaneously by combining in situ perfusion with in vitro perfusion, density gradient centrifugation, and differential attachment. The cell yield rate, cell viability and purity all met requirements for the subsequent cell experiment. Conclusion The combined cell isolation and culture method is feasible to isolate primary rat hepatocytes, hepatic stellate cells, Kupffer's cells and hepatic sinus endothelial cells simultaneously.
