Ameliorating injury during preservation and isolation of human islets using the two-layer method with perfluorocarbon and UW solution.
This study assessed the effects of a two-layer method (TLM), using perfluorocarbon and UW solution, on the quality of human pancreata following storage and islet yield/function after isolation. In part A, TLM was applied immediately after procurement and the energetic profile was compared to a group treated with UW solution only (control) throughout 24-h storage. In part B, cadaveric human pancreata were procured and subjected to a TLM after cold storage in UW solution (TLM group) or UW solution (control group). Energetics, lipid peroxidation, and islet recovery/function were assessed after preservation at 4 degrees C. In part A, after 9-h storage, the energetic profile (ATP, ATP/ADP, energy charge) for the TLM group was superior to controls. In part B, TLM treatment resulted in consistently greater ATP, ATP/ADP, and energy charge values than with storage in UW solution alone (p < 0.05). UW treatment resulted in 40% greater peroxidative damage than in the TLM group (p < 0.05). Islet recovery and functional viability were 30-40% higher following TLM treatment (p < 0.05). These data support the hypothesis that islet viability and yields can be significantly improved using a brief period of TLM treatment following conventional UW storage; reduced energetic and oxidative stress are implicated as potential mechanisms.
Salehi
Payam
P
Surgical-Medical Research Institute, Department of Surgery, University of Alberta, Edmonton, Alberta, Canada.
Mirbolooki
Mohammadreza
M
Kin
Tatsuya
T
Tsujimura
Toshiaki
T
Shapiro
A M James
AM
Churchill
Thomas A
TA
Lakey
Jonathan R T
JR
D000241:Adenosine; D000244:Adenosine Diphosphate / Q000032:analysis; D000255:Adenosine Triphosphate / Q000032:analysis; D000493:Allopurinol; D002469:Cell Separation; D002470:Cell Survival / Q000502:physiology; D003080:Cold Temperature; D015925:Cryopreservation / Q000379:methods*; D004734:Energy Metabolism / Q000502:physiology; D005466:Fluorocarbons; D005978:Glutathione; D006801:Humans; D007328:Insulin; D007515:Islets of Langerhans / Q000166:cytology / Q000293:injuries* / Q000502:physiology; D016381:Islets of Langerhans Transplantation / Q000379:methods*; D015227:Lipid Peroxidation / Q000502:physiology; D019188:Organ Preservation Solutions; D018384:Oxidative Stress / Q000502:physiology; D011309:Preservation, Biological / Q000379:methods; D011887:Raffinose; D013997:Time Factors
D005466:Fluorocarbons; D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D000244:Adenosine Diphosphate; D000493:Allopurinol; D000255:Adenosine Triphosphate; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose