Improved human islet isolation outcome from marginal donors following addition of oxygenated perfluorocarbon to the cold-storage solution

Abstract

Last year, from the approximately 6,000 organ donors, only approximately 1,500 pancreata were used for clinical transplantation. Factors that contribute to this poor pancreas use include strict donor selection criteria and the requirement for short cold-ischemia time (CIT). Numerous pancreata have not been used because of long ischemia times postprocurement. Given the oxygen-rich environment of the islets in the native pancreas, it is conceivable that islets are highly susceptible to irreversible damage following prolonged ischemia. The use of continuously oxygenated perfluorohydrocarbons (PFCs), known for their high oxygen-solubility coefficients, in a two-layer culture with standard University of Wisconsin preservation media, has extended the acceptable range CIT, and, furthermore, there has been no evidence of adverse effects from PFCs on the outcome of transplanted cells, whereas they often enhance islet cell function. The purpose of this study was to use the two-layer culture method to improve donor-organ use from marginal donors. Fifteen organs were procured using the two-layer method, and 18 without using it, from donors greater than 50 years of age. Despite nonsignificant differences in age, weight of the donors, weight of the organ and CIT, the PFC group yielded an average of twofold more islet equivalents than those harvested from the control group. As a result, from the control group, only 2 of 18 organs were used for clinical islet transplantation, whereas 8 of 15 were used from the PFC group. To this end, the two-layer method may help clinicians overcome the problem of organ underuse.

Technical aspects of islet preparation and transplantation

“…We have demonstrated that pancreata preserved in UW solution for prolonged periods ( > 10 h) can be rescued by an additional 3 h of preservation with the TLM [226,227,228,2291. The TLM had a positive effect on in vitro insulin secretory activity as compared to cold storage in UW solution alone [228,2291.…”

Polyphenol, an Extract of Green Tea, Increases Culture Recovery Rates of Isolated Islets from Nonhuman Primate Pancreata and Marginal Grade Human Pancreata

“…However, it is reported that islet mass is tients (19)(20)(21). Recent clinical trials carried out in different centers have definitively shown that islet transplants often lost due to apoptosis during isolation, culturing, and transplantation (6), which can account for up to can result in insulin independence (4,17). Among the factors contributing to this relatively recent success are a 60% loss of β-cells with transplantation (2).…”

“…The distended pancreas Viability was determined using fluorescein diacetate and propidium iodide (FDA/PI) (Molecular Probes, Eugene, was cut into 7-9 pieces and transferred to a Ricordi chamber (19). Digestion of the pancreas was facilitated OR) (17,18). The islets were scored for shape (flat vs. spherical), border (irregular vs. well rounded), integrity by recirculating enzyme solution through the Ricordi chamber at 37°C and shaking the chamber.…”

Chemical Oxygenation of Pancreatic Tissue Prior to Islet Isolation and Transplantation

“…These promising results could be confirmed in a similar setting by achieving a significantly higher islet transplantation rate of 71% in pancreases that were oxygenated for additionally 3 hours compared to UWS storage alone (36%) (Tsujimura et al, 2002a). An even more impressing improvement of the success rate from 11% to 53% was obtained in TLM-oxygenated pancreases from marginal donors older than 50 years (Ricordi et al, 2003). However, it should be stressed that in 80% of the organs used in the latter study oxygenation started immediately after retrieval utilizing oxygen-precharged PFD for oxygen supply.…”