Research Article Solubilized Pancreatic Extracellular Matrix from Juvenile Pigs ProtectsIsolatedHumanIsletsfromHypoxia-InducedDamage:A Viable Option for Clinical Islet Transplantation Heide Brandhorst, 1 Stasia Krishtul, 2 Daniel Brandhorst , 1 Limor Baruch, 2 Marcelle Machluf, 2 and Paul R. V. Johnson 1,3 1 Nufeld Department of Surgical Sciences, University of Oxford, Oxford No. 3 9DU, UK 2 Laboratory for Cancer Drug Delivery and Cell Based Technologies, Faculty of Biotechnology and Food Engineering, Technion Israel Institute of Technology, Haifa, Israel 3 Oxford Biomedical Research Centre (OxBRC), Oxford No. 3 9DU, UK CorrespondenceshouldbeaddressedtoDanielBrandhorst;daniel.brandhorst@nds.ox.ac.uk Received 26 January 2023; Revised 26 May 2023; Accepted 7 June 2023; Published 11 July 2023 AcademicEditor:CherieL.Stabler Copyright©2023HeideBrandhorstetal.TisisanopenaccessarticledistributedundertheCreativeCommonsAttribution License,whichpermitsunrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalworkisproperly cited. Tepancreaticextracellularmatrix(ECM)isanenormouslycomplexconstruct.Previousstudiesunderlinethechallengestoidentifythe optimalcombinationsandratiosofindividualECMproteinsforpromotingsurvivalandfunctionofisolatedandtransplantedislets.Tis studyaimedonassessingtheefciencyofsolubilizednaturalECMextractedfromjuvenilepigs,anunlimiteddonorsource.Isolated humanisletswereculturedunderahypoxicatmosphere(2%oxygen)inmediasupplementedwitheithersolubilizedporcinepancreatic ECM(ppECM)oramixtureofhumanECMproteinscomposedofcollagen•IV,laminin•521,andnidogen•1(hEPM).Controlislets wereculturedunderidenticalconditionswithoutECM•compounds.Reactiveoxygenspeciesproductionincreasedthree•foldincontrols butwasreducedbyhEPMorppECM.EarlyapoptosisremainedonpreculturelevelswhenisletsweretreatedwithhEPMorppECM. PrecultureviabilitywaspreservedwhenhEPMorppECMwasadministered.Whilstcontrolsfailedtorespondtoglucosechallenge, treatmentwithhEPMorppECMpreservedthephysiologicalinsulinresponse.Insummary,overallsurvivalwassignifcantlyhighestin ppECM•treatedislets.Tisstudypresentsanewapproachtoprotecthumanisletsfromhypoxia•induceddamagebysupplementing mediawithppECMextractedfromanunlimiteddonorsource.Te fndingsmayalsoserveasstartingpoint foranovelencapsulation technique to protect isolated human islets. 1.Introduction Te requirement to extract a subpopulation of heteroge• neous cell clusters from an organ whilst preserving their nativestructureandorganizationisauniquechallengefaced duringpancreaticisletisolationcomparedwithothertypes of tissue separation [1]. Tis requires a combination of enzymaticandmechanicaltechniques.Duringthisprocess, thepancreaticmatrixisdigested,andtheisletsreleasedfrom thedispersedacinartissuebeforethetwotissuesaresepa• ratedbydensity•gradientpurifcation.Akeyaspectofislet extraction, therefore, is the enzymatic detachment or cleavage of the islet basement membrane from the sur• rounding pancreatic matrix [2]. As a consequence, the bi• directionalcommunicationbetweentheintegrinsexpressed ontheisletcellsurfaceandthebasementmembraneislost [3–6]. Tis, in turn, interrupts the signaling of vital physi• ologicalfunctionsfromtheproximalmicroenvironmentto theisletcellssufering fromincreasedratesofapoptosisand severe loss during islet culture [7, 8]. Te absence of an extracellular matrix (ECM) [9, 10], in combination with hypoxicconditions[11]andashortageofessentialnutrients [12], is the key factor resulting in 73% of islet recipients currently requiring two or more islet grafts to achieve Hindawi Journal of Tissue Engineering and Regenerative Medicine Volume 2023, Article ID 7452682, 10 pages https://doi.org/10.1155/2023/7452682