Analysis of donor factors affecting human islet isolation with current isolation protocol.
BACKGROUND
Appropriate donor selection is one of the keys for successful human islet isolation. Previous studies identified several critical donor factors; however, significant improvements in current human islet isolation protocols make reevaluation of donor factors necessary.


METHODS
Review was performed on 31 human islet isolations. Islet isolations were conducted using the standard automated islet isolation method with three protocol revisions that included the two-layer method (TLM) of pancreas preservation prior to islet isolation, usage of purified collagenase mixture Liberase, and continuous density gradient for islet purification. Factors leading to successful isolations (islet yield > 100,000 IE and static incubation stimulation index greater than 2.0) were analyzed. The impacts of various risk factors were also examined.


RESULTS
Donors in the successful islet isolation group had a significantly lower incidence of elevated peak transaminases and creatinine levels, lower usage of norepinephrine or cardiac arrest, less prolonged hospitalization (> 96 hours), and less prolonged preservation time of donor pancreata (>25 hours). The TLM extended acceptable preservation time of donor pancreata from 8 to 25 hours. When donors had no risk factor, the success rate was 14/16 (87.5%). In sharp contrast, when donors had two or more risk factors, the success rate was 0/7 (0%; P <.001).


CONCLUSIONS
Risk factors for human islet isolation with the current islet isolation protocol were identified. The decision to process pancreata based on review of donor factors should improve the consistency of human islet isolations and transplantation for curing type 1 diabetes.
Matsumoto
S
S
Islet and Cell Processing Laboratory, Puget Sound Blood Center/Northwest Tissue Center, University of Washington Department of Surgery, USA. shinichi@kuhp.kyoto-u.ac.jp
Zhang
G
G
Qualley
S
S
Clever
J
J
Tombrello
Y
Y
Strong
D M
DM
Reems
J A
JA
D000328:Adult; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods; D017364:Collagenases; D005260:Female; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology*; D008297:Male; D015203:Reproducibility of Results; D012189:Retrospective Studies; D014019:Tissue Donors / Q000706:statistics & numerical data*
D017364:Collagenases