Investigation on the function and morphometrics of cultured primary rat ~hepatocytes
- VernacularTitle:长期培养的大鼠原代肝细胞功能和形态学观察
- Author:
Peng ZHUANG
;
Yuansen JIANG
;
Huihui MA
;
Zhigang LI
;
Li MAI
;
Lin YANG
;
Jilu YAO
- Publication Type:Journal Article
- Keywords:
Hepatocytes;
Cell culture
- From:
Chinese Journal of Pathophysiology
1989;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To investigate the function and morphological changes of long-term cultured primary rat hepatocytes. METHODS: Rat primary hepatocytes were isolated by two-step in situ collagenase perfusion method, and then were purified by density and grade centrifugal method with Percoll. Cell viability was observed by 0.4% trypan blue. The hepatocytes were seeded into 6 wells plate with HepatoZYME-SFM medium. ALT, AST, albumin and urea levels in the supernatant were measured, CYPⅠA1 was detected with EROD method. RESULTS: (2-3)?108 cells per whole liver were obtained with viability and purity above 90% after purified with Percoll. Hepatocytes cultured in HepatoZYME-SFM grew well with normal hepatocyte morphometrics. ALT, AST levels in the supernatant decreased after 3-day culture, and kept at a stable level after 6-9 days. Albumin secretion and urea synthesis were maintained at high levels in 18 days, while CYPⅠA 1 enzyme activity was only detected in 3-6 days. CONCLUSIONS: Percoll was used to increase the viability and purity of freshly isolated rat hepatocytes. Hepatocyte morphometrics and their biological synthesized function are effectively maintained in HepatoZYME-SFM medium.
