Caspase-3 inhibitor prevents apoptosis of human islets immediately after isolation and improves islet graft function.
OBJECTIVE
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 micromol/L Z-DEVD-FMK, 0.5% HSA + 100 micromol/L Z-DEVD-FMK and 10% FBS for 2 days. Next, 1000 IEQ islets precultured with 0.5% HSA and with or without 100 micromol/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.


CONCLUSIONS
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.
Nakano
Masahiko
M
Diabetes Institute for Immunology and Transplantation, Department of Surgery, University of Minnesota, Minneapolis, Minnesota 55455, USA.
Matsumoto
Ippei
I
Sawada
Toshiya
T
Ansite
Jeff
J
Oberbroeckling
Jeremy
J
Zhang
Hui Jian
HJ
Kirchhof
Nicole
N
Shearer
Jeff
J
Sutherland
David E R
DE
Hering
Bernhard J
BJ
D000328:Adult; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001786:Blood Glucose / Q000032:analysis; D002417:Cattle; D002469:Cell Separation; D002478:Cells, Cultured / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use; D003921:Diabetes Mellitus, Experimental / Q000601:surgery*; D004353:Drug Evaluation, Preclinical; D005260:Female; D005312:Fetal Blood; D006801:Humans; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D016381:Islets of Langerhans Transplantation; D007668:Kidney; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008875:Middle Aged; D009842:Oligopeptides / Q000494:pharmacology* / Q000627:therapeutic use; D012709:Serum Albumin; D013311:Streptozocin; D014183:Transplantation, Heterologous; D016042:Transplantation, Heterotopic
D001786:Blood Glucose; D003470:Culture Media; D015853:Cysteine Proteinase Inhibitors; D007328:Insulin; D009842:Oligopeptides; D012709:Serum Albumin; C110772:benzoylcarbonyl-aspartyl-glutamyl-valyl-aspartyl-fluoromethyl ketone; D013311:Streptozocin