Endothelialization of Gore-Tex vascular graft by using cryopreserved human umbilical endothelial cells
- VernacularTitle:人脐静脉内皮细胞分离与冻存技术及Gore-Tex人工血管内皮化
- Author:
Hongjun JIANG
;
Bing JIA
;
Zhanggen CHEN
- Publication Type:Journal Article
- Keywords:
Endothelium, vascular Cell culture Polytetrafluoroethylene Flow cytometry
- From:
Chinese Journal of Thoracic and Cardiovascular Surgery
1995;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective The feasibility of constructing endothelialized vascular graft by using cryopreserved HUVCs was studies. Methods Forty-two human umbilical cords were used in this study. HUVECs were isolated by means of filling umbilical veins with digestive enzyme solution. HUVECs were then cultured and observed. Endothelial cells were identified by von Willebrand factor immunofluorescence staining and scanning electron microscope examination. Endothelial cells were suspended in cryopreserving solution which contains 10% DMSO and 10% fetal bovine serum in M199 and cryopreserved in liquid nitrogen. Post-thawed cells and non-frozen cells proliferation was evaluated by measuring the metabolic activity of tetrazolium compound. The endothelial cell growth characteristics were determined by daily observation using phase contrast microscope. Post-thawed endothelial cells viability was determined by trypan blue staining test. Flow cytometry were applied to determine the apoptosis rate of post-thawed cells. Cryopreserved endothelial cells morphological examinations such as hematoxylin and eosins staining and scanning electron microscope examination were carried out in this study. After cell culture and amplification, cryopreserved HUVCs were seeded on the inner surface of Gore-Tex graft to construct tissue engineered vascular graft. Results Extreme high-purified endothelial cells could be isolated by infusing digestive solutions to the lumen of human umbilical veins. Compared with non-frozen endothelial cells, Post-thawed endothelial cells showed 95% of vitality. Post-thawed HUVEC growth curve was similar to non-frozen ones'. Post-thawed HUVEC apoptosis rate (5.85? 0.56) % was higher than non-frozen ones (5.34?0.49)%; however, the difference was not statistically different. Endothelialization of vascular graft was carried out successfully. Cryopreserved cells on Gore-Tex surface showed a good growth trend. Conclusion Cryopreserved HUVCs may be taken as a cell choice for tissue engineering. Enough high pure endothelial cells could be isolated by digestive solutions infusion of human umbilical veins. Post-thawed endothelial cells are proved to have high vitality and growth potential on Gore-Tex surface in vitro.
