Lower beta cell yield from donor pancreases after controlled circulatory death prevented by shortening acirculatory warm ischemia time and by using IGL-1 cold preservation solution

Abstract

Organs from donors after controlled circulatory death (DCD III) exhibit a higher risk for graft dysfunction due to an initial period of warm ischemia. This procurement condition can also affect the yield of beta cells in islet isolates from donor pancreases, and hence their use for transplantation. The present study uses data collected and generated by our Beta Cell Bank to compare the number of beta cells in isolates from DCD III (n = 141) with that from donors after brain death (DBD, n = 609), before and after culture, and examines the influence of donor and procurement variables. Beta cell number per DCD III-organ was significantly lower (58 x 106 versus 84 x 106 beta cells per DBD-organ; p < 0.001) but their purity (24% insulin positive cells) and insulin content (17 μg / 106 beta cells in DCD III-organs versus 19 μg / 106 beta cells in DBD-organs) were similar. Beta cell number correlated negatively with duration of acirculatory warm ischemia time above 10 min; for shorter acirculatory warm ischemia time, DCD III-organs did not exhibit a lower beta cell yield (74 x 106 beta cells). Use of Institut Georges Lopez-1 cold preservation solution instead of University of Wisconsin solution or histidine-tryptophan-ketoglutarate also protected against the loss in beta cell yield from DCD III-organs (86 x 106 for IGL-1 versus 54 x 106 and 65 x 106 beta cells respectively, p = 0.042). Multivariate analysis indicates that both limitation of acirculatory warm ischemia time and use of IGL-1 prevent the reduced beta cell yield in islet cell isolates from DCD III-organs.

Hypothermic oxygenated machine perfusion of the human pancreas for clinical islet isolation: a prospective feasibility study

“…In three small studies concerning islet isolation from DCD donors, no significant differences in islet yield obtained from DCD compared with DBD pancreases were reported [ 43 , 44 , 45 ]. However, two more recent studies reported a reduction of up to 100 000 IEQ from DCD pancreases compared with DBD pancreases [ 10 , 11 ], and another study reporting a 30% lower yield for DCD pancreases when calculated as beta‐cell number [ 46 ].…”

Utility of Islet Cell Preparations From Donor Pancreases After Euthanasia

“…A rapid sterno-laparotomy is then performed followed by cold flush, topical cooling, and procurement of the pancreas. Because of its beneficial effect on DCD pancreas preservation for islet cell isolation 13 , IGL-1 is used as cold preservation solution in our DCD V procurement. Total WIT is defined as time between end-of-life therapy and abdominal aorta cannulation initiating cold preservation.…”

“…Between January 2015 and December 2020, 13 DCD V organs that matched our set criteria were processed and included for analysis. They were compared with our reference cohort of 234 donors after brain death (DBD) from the same time period and a cohort of 29 DCD III (donors after controlled circulatory death) organs selected for acirculatory WIT <10 min and IGL-1 preservation solution, conditions that have been shown to favor DCD islet cell isolation 13 . Reasons for euthanasia request are shown in Table 1 .…”

“…Our university hospital is strongly involved in end-of-life care with referrals specific for these procedures. This allowed us to evaluate in vitro and in vivo outcome of islet cell isolations from these donors while selecting conditions that have been shown beneficial for DCD organ procurement and preservation, such as the use of Institut Georges Lopez-1 (IGL-1) preservation solution and limiting acirculatory warm ischemia to 10 min 13 .…”

Function and composition of pancreatic islet cell implants in omentum of type 1 diabetes patients

“…Previous work has shown that induction of plasma Cpeptide-positivity required minimally 2.10 6 cultured beta cells/ kg recipient bodyweight (BW) at minimally 20 percent purity. 2 We choose beta cell number instead of islet equivalents (IEQ) because it directly and specifically expresses beta cell mass while dithizonestaining for IEQ measurement is not beta cell-specific and weakens following culture; the corresponding IEQ can be estimated with a conversion factor of 450 beta cells/IEQ which is taken from prior studies where both parameters were measured 7,8 (Table 1). Donors were ABO-compatible and passed a leucocyte crossmatch but were not human leucocyte antigen (HLA)-matched; they were negative for HIV, hepatitis B and C.…”