Caspase-3 Inhibitor Prevents Apoptosis of Human Islets... : Pancreas (original) (raw)

Original Article

Caspase-3 Inhibitor Prevents Apoptosis of Human Islets Immediately After Isolation and Improves Islet Graft Function

Nakano, Masahiko MD; Matsumoto, Ippei MD; Sawada, Toshiya MD; Ansite, Jeff MD; Oberbroeckling, Jeremy; Zhang, Hui Jian MD; Kirchhof, Nicole Veterinary, MD; Shearer, Jeff; Sutherland, David E. R. MD; Hering, Bernhard J. MD

From the Diabetes Institute for Immunology and Transplantation, Department of Surgery, University of Minnesota, Minneapolis, Minnesota.

Received for publication September 2, 2003; accepted February 17, 2004.

Reprints: Bernhard J Hering, MD, Diabetes Institute for Immunology and Transplantation Department of Surgery, University of Minnesota, 420 Delaware St. SE, Minneapolis, MN 55455 (e-mail: [email protected]).

Abstract

Objectives:

Apoptosis appears in islets after isolation, and it has a detrimental effect on the islet function. To improve the outcome of clinical islet transplantation, it is crucial to protect islets from apoptosis. The aim of this study was to determine whether a caspase-3 inhibitor (Z-DEVD-FMK) added to culture media protects islets from apoptosis and to compare the effects of fetal bovine serum (FBS) with human serum albumin (HSA) as a protein supplement in culture.

Methods:

Isolated human islets were cultured under 4 different conditions: 0.5% HSA (control), 0.5% HSA + 25 μmol/L Z-DEVD-FMK, 0.5% HSA + 100 μmol/L Z-DEVD-FMK and 10% FBS for 2 days. Next, 1000 IEQ islets precultured with 0.5% HSA and with or without 100 μmol/L Z-DEVD-FMK were transplanted to diabetic nude mice.

Results:

The islet yields were higher in Z-DEVD-FMK–treated groups, and the inhibitor prevented apoptosis dose dependently. The yield and insulin release were higher in FBS-treated group than in the control group, but FBS did not affect apoptosis. All 6 mice transplanted with islets pretreated with Z-DEVD-FMK, and 3 of 8 mice with control islets became normoglycemic posttransplantation.

Conclusion:

Z-DEVD-FMK prevented apoptosis of isolated human islets and improved its function. FBS (10%) improved the islet yield and insulin secretion more than 0.5% HSA.