Isolation and purification of pancreatic islets in adult donors
10.3760/cma.j.issn.1674-6090.2011.05.003
- VernacularTitle:成人胰岛细胞的分离与纯化
- Author:
Jianxin JIANG
;
Chengyi SUN
- Publication Type:Journal Article
- Keywords:
Pancreatic islet;
Cell isolation;
Cell purification
- From:
Journal of Endocrine Surgery
2011;05(5):296-298
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the methods and feasibility of islets isolation and purification,and to get more islets with high purity and good function for clinical transplantation.Methods After being weighted,human pancreatic islets were isolated with type Ⅴ collagenase,and purified by Ficoll's discontinuous density gradient centrifugation.The yield and purity of islets were evaluated by dithizone(DTZ) staining under microscope,and the viability was assessed by insulin release assay in vitro.Results The average number of islets was about 3 600 ± 447 per gram pancreas after isolation and it was about 2140 ±207 after purification with more than 70% purity.2,4,6 days after islet cell culture,the basal insulin concentration of the culture medium was measured,and it was 3.302 ± 1.63,3.504 ±1.10,and 2.921 ±1.13 (mIU/L/100 islets) respectively.Conclusion Collagenase digest and Ficoll's discontinuous density gradient centrifugation are effective methods for isolation and purification of human pancreatic islets.