Culture of cryopreservted hepatocytes of sucking pig with collagen gel
- VernacularTitle:冻存乳猪肝细胞的胶原凝胶培养研究
- Author:
Hongling LIU
;
Huifen WANG
;
Yingjie WANG
- Publication Type:Journal Article
- Keywords:
hepatocytes;
cell culture;
cryopreservation;
collagen
- From:
Medical Journal of Chinese People's Liberation Army
2001;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the method of cryopreservation, and method of cultivation of hepatocytes of sucking pig. Methods Hepatocytes were isolated from sucking pig by two-step perfusion with collagenase using an extracorporeal perfusion apparatus. They were cryopreserved in liquid nitrogen for 4 months. They were thawed and seeded on gelatin coated bottom of a flask. The cells were covered with another layer of collagen. The exprssion of albumin mRNA, cellular morphology synthesis of urea, and content of oxalylacetylaminotransferase were determined in the collagen cultures. Results Thawed hepatocytes maintained their biological characters well after cryopreservation. The thawed hepatocytes could grow well in collagen gel, sandwich culture. In the 10-day cultures, typical cellular morphological and biological characteristics of cultured hepatocytes could be observed. Conclusion Hepatocytes stored in liquid nitrogen could maintain their activities for a long time. The hepatocytes could be cultivated in collagenous matrix, which provided an environment closely resembling that in nature, and their morphology and functions could be maintained.
