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Beneficial effect of recombinant rC1rC2 collagenases on human islet function: Efficacy of low dose enzymes on pancreas digestion and yield.

AMERICAN JOURNAL OF TRANSPLANTATION(2018)

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摘要
A high number of human islets can be isolated by using modern purified tissue dissociation enzymes; however, this requires the use of >20 Wunsch units (WU)/g of pancreas for digestion. Attempts to reduce this dose have resulted in pancreas underdigestion and poor islet recovery but improved islet function. In this study, we achieved a high number of functional islets using a low dose of recombinant collagenase enzyme mixture (RCEM-1200 WU rC2 and 10 million collagen-degrading activity [CDA] U of rC1 containing about 209mg of collagenase to digest a 100-g pancreas). The collagenase dose used in these isolations is about 42% of the natural collagenase enzyme mixture (NCEM) dose commonly used to digest a 100-g pancreas. Low-dose RCEM was efficient in digesting entire pancreases to obtain higher yield (5535830 and 2582 +/- 925 islet equivalent/g, P<.05) and less undigested tissue (16.7 +/- 5% and 37.8 +/- 3%, P<.05) compared with low-dose NCEM (12WU/g). Additionally, low-dose RCEM islets retained better morphology (confirmed with scanning electron microscopy) and higher in vitro basal insulin release (2391 +/- 1342 and 1778 +/- 978 U/mL; P<.05) compared with standard-dose NCEM. Nude mouse bioassay demonstrated better islet function for low-dose RCEM (area under the curve [AUC] 24968) compared with low-dose (AUC-38225) or standard-dose NCEM (AUC-38685), P<.05. This is the first report indicating that islet function can be improved by using low-dose rC1rC2 (RCEM). This study evaluates the efficacy of low-dose recombinant collagenase enzyme mixture and identifies its beneficial effect on human islet yield and function.
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关键词
islet isolation,islet transplantation,islets of Langerhans,translational research,science
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