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

Masafumi Goto, Torsten M. Eich, Marie Felldin, Aksel Foss, Ragnar Källen, Kaija Salmela, Annika Tibell, Gunnar Tufveson, Keisei Fujimori, Margareta Engkvist, Olle Korsgren

研究成果: Article査読

178 被引用数 (Scopus)

抄録

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.

本文言語English
ページ(範囲)1367-1375
ページ数9
ジャーナルTransplantation
78
9
DOI
出版ステータスPublished - 2004 11月 15
外部発表はい

ASJC Scopus subject areas

  • 移植

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