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Refinement of the automated method for human islet isolation and presentation of a closed system for in vitro islet culture

Goto, M; Eich, TM; Felldin, M; Foss, A; Källén, Ragnar LU ; Salmela, K; Tibell, A; Tufveson, G; Fujimori, K and Engkvist, M, et al. (2004) In Transplantation 78(9). p.1367-1375
Abstract
Background. The procedure of human islet isolation needs further optimization and standardization. Here, we describe techniques to enhance enzymatic digestion and minimize mechanical forces during the digestion process. The isolation protocol has also been modified to meet current GMP (cGMP) standards. Moreover, the impact of donor- and process-related factors was correlated to the use of islets for clinical transplantation. Methods. One hundred twelve standardized consecutive islet isolations were evaluated. Metyltioninklorid and indermil (topical tissue adhesive) were applied to detect leakage of collagenase injected and to repair the damaged pancreatic glands. The effects of dye and glue were evaluated in terms of islet yield, islet... (More)
Background. The procedure of human islet isolation needs further optimization and standardization. Here, we describe techniques to enhance enzymatic digestion and minimize mechanical forces during the digestion process. The isolation protocol has also been modified to meet current GMP (cGMP) standards. Moreover, the impact of donor- and process-related factors was correlated to the use of islets for clinical transplantation. Methods. One hundred twelve standardized consecutive islet isolations were evaluated. Metyltioninklorid and indermil (topical tissue adhesive) were applied to detect leakage of collagenase injected and to repair the damaged pancreatic glands. The effects of dye and glue were evaluated in terms of islet yield, islet function using the perifusion assay, and success rate of the isolation. To analyze key factors for successful isolations, both univariate and multivariate regression analysis were performed. Results. Both Metyltioninklorid and Indermil were effective to prevent leakage of enzyme solutions from the pancreatic glands. Both islet yield and success rate were higher when these tools were applied (4,516.1 +/- 543.0 vs. 3,447.7 +/- 323.5, P=0.02; 50.0% vs. 21.3%, P=0.02, respectively). No adverse effects on islet function or collagenase activity were observed. Multivariate regression analysis identified the maximal recorded amylase >100 U/L (P=0.026), BMI (P=0.03), and the use of catecholamine (P=0.04) as crucial donor-related factors. In addition, cold ischemia time (P=0.005), the dissection procedure using whole glands with duodenum (P=0.02), and the local procurement team (P=0.03) were identified as crucial isolation-related variables. Conclusions. A standardized technique of islet isolation is presented applying novel means to improve enzymatic digestion and to meet cGMP standards. (Less)
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published
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keywords
human islet isolation, tissue glue, dye, dissection procedure, culture bag system, multivariate statistical analysis
in
Transplantation
volume
78
issue
9
pages
1367 - 1375
publisher
Lippincott Williams & Wilkins
external identifiers
  • wos:000225192700020
  • pmid:15548977
  • scopus:8644282710
ISSN
1534-6080
language
English
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yes
id
566437ff-6863-4227-b5ca-8703967e4740 (old id 897857)
alternative location
http://www.transplantjournal.com/pt/re/transplantation/abstract.00007890-200411150-00020.htm
date added to LUP
2008-01-10 15:19:16
date last changed
2017-12-17 03:53:32
@article{566437ff-6863-4227-b5ca-8703967e4740,
  abstract     = {Background. The procedure of human islet isolation needs further optimization and standardization. Here, we describe techniques to enhance enzymatic digestion and minimize mechanical forces during the digestion process. The isolation protocol has also been modified to meet current GMP (cGMP) standards. Moreover, the impact of donor- and process-related factors was correlated to the use of islets for clinical transplantation. Methods. One hundred twelve standardized consecutive islet isolations were evaluated. Metyltioninklorid and indermil (topical tissue adhesive) were applied to detect leakage of collagenase injected and to repair the damaged pancreatic glands. The effects of dye and glue were evaluated in terms of islet yield, islet function using the perifusion assay, and success rate of the isolation. To analyze key factors for successful isolations, both univariate and multivariate regression analysis were performed. Results. Both Metyltioninklorid and Indermil were effective to prevent leakage of enzyme solutions from the pancreatic glands. Both islet yield and success rate were higher when these tools were applied (4,516.1 +/- 543.0 vs. 3,447.7 +/- 323.5, P=0.02; 50.0% vs. 21.3%, P=0.02, respectively). No adverse effects on islet function or collagenase activity were observed. Multivariate regression analysis identified the maximal recorded amylase >100 U/L (P=0.026), BMI (P=0.03), and the use of catecholamine (P=0.04) as crucial donor-related factors. In addition, cold ischemia time (P=0.005), the dissection procedure using whole glands with duodenum (P=0.02), and the local procurement team (P=0.03) were identified as crucial isolation-related variables. Conclusions. A standardized technique of islet isolation is presented applying novel means to improve enzymatic digestion and to meet cGMP standards.},
  author       = {Goto, M and Eich, TM and Felldin, M and Foss, A and Källén, Ragnar and Salmela, K and Tibell, A and Tufveson, G and Fujimori, K and Engkvist, M and Korsgren, O},
  issn         = {1534-6080},
  keyword      = {human islet isolation,tissue glue,dye,dissection procedure,culture bag system,multivariate statistical analysis},
  language     = {eng},
  number       = {9},
  pages        = {1367--1375},
  publisher    = {Lippincott Williams & Wilkins},
  series       = {Transplantation},
  title        = {Refinement of the automated method for human islet isolation and presentation of a closed system for in vitro islet culture},
  volume       = {78},
  year         = {2004},
}