The performance of different isolation regents for purification of mice primary hepatic stellate cells
10.3760/cma.j.issn.1003-9279.2014.01.022
- VernacularTitle:不同分离介质对小鼠原代肝星状细胞分离纯化效果的影响
- Author:
Hongmin LI
1
;
Hui REN
;
Xiaohua HAO
;
Yubo HUANG
;
Hongshan WEI
Author Information
1. 首都医科大学附属北京地坛医院
- Keywords:
Liver cirrhosis;
Cells,cultured;
Cells/isolation and purification;
Mice
- From:
Chinese Journal of Experimental and Clinical Virology
2014;28(1):64-66
- CountryChina
- Language:Chinese
-
Abstract:
Objective To compare the advantages and disadvantages of the three isolation regents for isolating primary hepatic stellate cells(HSCs) of mice.Methods The mice HSCs was isolated by in situ perfusion of per-perfusion solution,perfusion with collagenase Ⅳ.After digestion,Ficoll-Paque PLUS,perco11 and Nycodenz were use for the gradient centrifugation.To calculate the quantity,purity and the viability of the HSCs.The autofluorescence and Oil O staining were used to identify primary HSCs.Results The quantity of HSCs was (1.0 ± 0.5) × 106,(1.0 ± 0.5) × 105,(0.8 ± 0.5) × 106/ g_liver,respectively.Using Ficoll-Paque PLUS,percoll and Nycodenz,the purity of primary HSCs were 75％-85％,91％-95％,91％-95％,respectively.The viability were all above 90％.Conclusion compared with Ficoll-Paque PLUS and percoll,the Nycodenz is the the best separating medium for isolation and purification of mice primary HSCs.
