Urop.due.uci.edu
Effect of In Vitro Culture under Hypoxic Conditions on Young Porcine Islet Viability
Nick Neel
Mentor: Jonathan Robert Todd Lakey
In transplanting islets, hypoxic conditions are encountered due to the lack of vascularization surrounding the newly transplanted islets. In order to survive, transplanted islets need to be able to withstand hypoxic conditions during the immediate post-transplant period until they become vascularized. We analyzed the effects of in vitro culture in various oxygen concentrations on islet viability. 10,000 IEq of 18-24 day old YP islets were cultured for seven days in vitro in UCI maturation media. 5000 IEq of islets were encapsulated in UP LVM alginate, while 5000 IEq were left unencapsulated. Both groups of islets were then incubated in a hypoxia chamber at 0%, 10% and 20% O2 for 72 hours. After 72 hours, 200 IEq of islets were stained with standard viability (Calcein Blue/Propidium Iodide) and apoptosis stains (YoPro-1). The islets were examined for viability after twenty-four and seventy-two hours. Islets incubated at 20% O2 demonstrated greater viability than those incubated under hypoxic conditions. Islet viability fell drastically between 24 and 72 hours of hypoxic culture. No significant difference in viability was noted between the free and encapsulated islet groups. The results demonstrate that free and encapsulated islets are not able to survive in hypoxic conditions for prolonged periods of time. This indicates that successful encapsulated islet transplant can be achieved by strategies aimed to prevent tissue loss due to hypoxia in the immediate post-transplant period.
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