Improved outcome of pig islet isolation by pefabloc inhibition of trypsin

Basir, Ibrahim; Burg, M.P.M. van der; Scheringa, Marcel; Töns, Annemiek; Bouwman, E

doi:10.1016/s0041-1345(97)00168-1

Cited by 25 publications

(12 citation statements)

References 2 publications

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“…The addition of Pefabloc, Trasylol and UTI during the islet isolation process has been reported to improve islet yield, presumably by inhibiting pancreatic endogenous protease activity. [16][17][18][19][20][21][22][23][24][25][26][27][28] To confirm this, we tested the effect of these inhibitors on the activity of trypsin, chymotrypsin, elastase and switch samples. Pancreatic digestion switch samples usually consist of multiple enzymes (trypsin, chymotrypsin, elastase, thermolysin/neutral protease and collagenase).…”

Section: Resultsmentioning

confidence: 99%

“…15 It has been shown that addition of protease inhibitors, such as Pefabloc, Trasylol or Ulinastatin (UTI), during islet isolation, resulted in improved yield. [16][17][18][19][20][21][22][23] However, the effects of these protease inhibitors on endogenous proteases and enzymes essential for tissue dissociation, in particular neutral protease, have yet to be determined. This study investigated the effect of three protease inhibitors, Pefabloc, Trasylol and UTI, on pancreatic endogenous proteases, thermolysin/neutral protease and islet function.…”

Section: Introductionmentioning

confidence: 99%

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Serine protease inhibitors suppress pancreatic endogenous proteases and modulate bacterial neutral proteases

Nduaguibe¹,

Bentsi-Barnes²,

Mullen³

et al. 2010

Islets

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Serine protease inhibitors suppress pancreatic endogenous proteases and modulate bacterial neutral proteases

Nduaguibe¹,

Bentsi-Barnes²,

Mullen³

et al. 2010

Islets

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“…The protease inhibitor Pefabloc (Roche Molecular Biochemicals, Indianapolis, IN) was used in groups 2, 3, and 4 (Pefabloc, TLM π Pefabloc, and UW π Pefabloc, respectively). The Pefabloc concentration (4.0 mmol) was based on published reports (8)(9)(10) and the recommendation of the manufacturer.…”

Section: Macaca Nemestrina Islet Isolation With the Seattle Open-pan mentioning

confidence: 99%

“…Before islet isolation, we used two-layer method (TLM) preservation to oxygenate the pancreas (5), and during isolation we inhibited trypsin activity with the serine-protease inhibitor Pefabloc [AEBSF; 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride]. Separately, both pancreas TLM preservation (6,7) and Pefabloc have increased islet yields (8)(9)(10)(11)(12) in both animals and in humans.…”

Section: Introductionmentioning

confidence: 99%

“…Pefabloc used during pig islet isolation improved the poor islet yields that accompanied pancreas protease activation (8)(9)(10). We speculated that endogenous trypsin activity reduces islet yields by eroding the pancreatic ductules, creating leaks that compromise the crucial delivery of collagenase digestion solution throughout the pancreas (18).…”

Section: Introductionmentioning

confidence: 99%

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Improved Islet Yields From Macaca Nemestrina and Marginal Human Pancreata After Two‐Layer Method Preservation and Endogenous Trypsin Inhibition

Matsumoto

Rigley

Reems

et al. 2003

American Journal of Transplantation

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We tested whether two-layer method (TLM) pancreas preservation and trypsin inhibition (Pefabloc) during processing allows longer preservation while retaining or improving viable islet recovery. Non-marginal primate (Macaca nemestrina) and marginal human (ischemic or preservation-injured) pancreata were processed with a research-oriented pan technique (Seattle method). Organs were processed upon arrival (∫ Pefabloc), or after TLM or University of Wisconsin solution (UW) preservation (π Pefabloc). Islet yield, viability, and function were assessed. Pefabloc increased M. nemestrina islet yields from 9696 ∫ 1749 IE/g to 15822∫ 1332 IE/g (p ∞ 0.01). Twolayer method preservation (∞ 6 h) further increased yields, to 23 769 ∫ 2773 IE/g (vs. π Pefabloc; p ∞0.01). Similarly, Pefabloc increased marginal human islet yields from 2473 ∫ 472 IE/g to 4723∫ 1006 IE/g (p ∞ 0.04). This increase was maintained after lengthy TLM preservation (Ͼ 30 h; 4801 ∫ 1066 IE/g). We also tested the applicability of TLM preservation (23.5 ∫ 3.2 h) to the processing of marginal human pancreata by the Edmonton/Immune Tolerance Network clinical protocol. Islet yield and function approached published results of pancreata processed 4.8 ∫ 0.8 h after organ recovery (p Ω 0.06). Pefabloc, and TLM vs. UW preservation, prolonged the tolerable interval between organ recovery and islet isolation. Islet yield, viability, and functionality improved from both marginal and nonmarginal pancreata.

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Pancreatic Islets: Methods for Isolation and Purification of Juvenile and Adult Pig Islets

Brandhorst

Johnson

Brandhorst

2016

Advances in Experimental Medicine and Biology

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The current situation of organ transplantation is mainly determined by the disbalance between the number of available organs and the number of patients on the waiting list. This obvious dilemma might be solved by the transplantation of porcine organs into human patients. The metabolic similarities which exist between both species made pancreatic islets of Langerhans to that donor tissue which will be most likely transplanted in human recipients. Nevertheless, the successful isolation of significant yields of viable porcine islets is extremely difficult and requires extensive experiences in the field. This review is focussing on the technical challenges, pitfalls and particularities that are associated with the isolation of islets from juvenile and adult pigs considering donor variables that can affect porcine islet isolation outcome.

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Improved outcome of pig islet isolation by pefabloc inhibition of trypsin

Cited by 25 publications

References 2 publications

Serine protease inhibitors suppress pancreatic endogenous proteases and modulate bacterial neutral proteases

Serine protease inhibitors suppress pancreatic endogenous proteases and modulate bacterial neutral proteases

Improved Islet Yields From Macaca Nemestrina and Marginal Human Pancreata After Two‐Layer Method Preservation and Endogenous Trypsin Inhibition

Pancreatic Islets: Methods for Isolation and Purification of Juvenile and Adult Pig Islets

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