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

  1. Shinichi Matsumoto1,2,
  2. Theodore H. Rigley1,
  3. Jo Anna Reems1,3,
  4. Yoshikazu Kuroda4,
  5. R. Brian Stevens1,2,5,*

Article first published online: 19 DEC 2002

DOI: 10.1034/j.1600-6143.2003.30110.x

Issue

American Journal of Transplantation

American Journal of Transplantation

Volume 3, Issue 1, pages 53–63, January 2003

Additional Information

How to Cite

Matsumoto, S., Rigley, T. H., Reems, J. A., Kuroda, Y. and Stevens, R. B. (2003), Improved Islet Yields From Macaca Nemestrina and Marginal Human Pancreata After Two-Layer Method Preservation and Endogenous Trypsin Inhibition. American Journal of Transplantation, 3: 53–63. doi: 10.1034/j.1600-6143.2003.30110.x

Author Information

  1. 1

    Puget Sound Blood Center/North-west Tissue Center, Seattle, WA

  2. 2

    Department of Surgery, Division of Transplantation, University of Washington Medical Center, Seattle, WA

  3. 3

    Department of Hematology, Division of Medicine, University of Washington, Seattle, WA

  4. 4

    Department of Gastroenterological Surgery, Graduate School of Medical Sciences, Kobe University, Kobe, Japan

  5. 5

    University of Washington Diabetes Endocrinology Research Center, Seattle, WA

* *Correspondence: R. Brian Stevens, MD, PhD, rbstevens@surgery.unmc.edu

Publication History

  1. Issue published online: 19 DEC 2002
  2. Article first published online: 19 DEC 2002
  3. Received 21 May 2001,revised andaccepted for publication 31 January 2002

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Keywords:

  • Primate islet isolation (Macaca nemestrina);
  • human islet isolation;
  • trypsin inhibition;
  • Pefabloc;
  • two-layer method

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 (+ Pefabloc). Islet yield, viability, and function were assessed.

Pefabloc increased M. nemestrina islet yields from 9696 ± 1749 IE/g to 15 822 ± 1332 IE/g (p < 0.01). Two-layer method preservation (< 6 h) further increased yields, to 23 769 ± 2773 IE/g (vs. + 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 (> 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 = 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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