Study on the isolation,culture,cryopreservation,and thawing of rat hepatocyte in hepatocyte transplantation experimental research
- VernacularTitle:肝细胞移植实验研究中大鼠肝细胞的冻存与复苏初探
- Author:
Li LI
;
Gaojun TENG
- Publication Type:Journal Article
- Keywords:
Rats, Sprague Dawley;
Liver;
Cell separation;
Cell survival;
Cryopreservation;
Technology, medical laboratory;
Evaluation studies
- From:
Chinese Journal of Radiology
2001;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the effects of cryopreservation and two different cooling rates on rat hepatocytes. Methods Perfusion with collagenase digestion was used to isolate rat hepatocytes. Hepatocytes were incubated with a freezing medium consisting of RPMI 1640 with 20% calf serum and 10% DMSO. Two different freezing protocols were applied using a computer controlled freezer to freeze the medium to -80℃ before hepatocytes were plunged into liquid nitrogen and stored. The viability, morphology, and protein synthesis were measured at Day 3, Day 15, Day 30, respectively after thawing. Results Both viability and protein synthesis ability of group B and group C decreased significantly as compared with that of Group A ( P 0.05). Electron microscopy showed ultrastructural changes between fresh and thawed rat hepatocytes including injury of cell membrane, loss of cell content, alteration of nucleolus. The degree of injury in Group B is slighter than in Group C. Conclusion Cooling rate of cryopreservation correlates with the quality of thawed hepatocytes. The cryopreservation and thawing procedures are key factors to affect the quantity and quality of hepatocytes. The viability and protein synthesis ability are not influenced by the storage period in liquid nitrogen.
