MP29-11 REPOPULATION OF PRIMARY RENAL CELLS FOR WHOLE ORGAN ENGINEERING: FUNCTIONAL EVALUATIONS (2016)

You have accessJournal of UrologyTransplantation & Vascular Surgery: Renal Transplantation & Vascular Surgery I1 Apr 2016MP29-11 REPOPULATION OF PRIMARY RENAL CELLS FOR WHOLE ORGAN ENGINEERING: FUNCTIONAL EVALUATIONS Mehran Abolbashari, Mi-Kyung Lee, Sigrid Agcaoili, Tamer Aboushwareb, In Kap Ko, John Jackson, Anthony Atala, and James Yoo Mehran AbolbashariMehran Abolbashari More articles by this author , Mi-Kyung LeeMi-Kyung Lee More articles by this author , Sigrid AgcaoiliSigrid Agcaoili More articles by this author , Tamer AboushwarebTamer Aboushwareb More articles by this author , In Kap KoIn Kap Ko More articles by this author , John JacksonJohn Jackson More articles by this author , Anthony AtalaAnthony Atala More articles by this author , and James YooJames Yoo More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.1095AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Renal transplantation is the only definitive treatment for end stage renal disease (ESRD), however it is severely limited by donor shortage. Whole organ engineering based on decellularization/recellularization techniques using patient′s own cells has been proposed as an alternative approach to renal transplantation. We previously demonstrated that repopulation of primary renal cells within acellular porcine kidney scaffolds leads to cell attachment and organization with maintenance of cellular phenotypes. In this study, we examined whether the recellularized porcine kidney scaffolds possess functional capabilities. METHODS Primary porcine renal cells were expanded, characterized, and seeded into the cortex of decellularized renal scaffolds, followed by bioreactor perfusion culture for 14 days. To evaluate the renal function of the repopulated renal structures formed within the scaffold, the levels of erythropoietin (EPO) in culture media and EPO mRNA expression were determined. Electrolyte transport and albumin re-adsorption were evaluated by sodium and albumin uptake assays, respectively. The activity of amino acid transport gamma glutamyl transpeptidase (GGT) or leucine aminopeptidase (LAP) (hydrolase activity) on the renal cells was determined by measuring the transformed amino acids released in the cell culture medium as a product of hydrolysis of their substrates of L-glutamic acid p-nitroanilide or L-leucine p-nitroanilide, respectively. RESULTS Fluorescent microscopic results demonstrated the evidence of sodium and albumin uptake by the renal cells within the repopulated constructs. Hydrolase activity assay showed that the renal cells within the recellularized construct expressed a significant hydrolase activity, indicating that the re-organized renal structures within the recellularized kidney constructs retain similar levels of hydrolase activity as native kidneys. The results of EPO quantification showed that the seeded cells produced EPO and its levels increased over time in the media, which was also confirmed by sufficient amounts of EPO mRNA by the seeded cells. CONCLUSIONS These functional evaluations indicate that the repopulated cells within the kidney construct show renal functions such as re-adsorption of sodium and protein, hydrolase activity, and production of erythropoietin. These results provide preliminary data for the generation of a transplantable renal graft as a potential treatment for ESRD. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e384 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Mehran Abolbashari More articles by this author Mi-Kyung Lee More articles by this author Sigrid Agcaoili More articles by this author Tamer Aboushwareb More articles by this author In Kap Ko More articles by this author John Jackson More articles by this author Anthony Atala More articles by this author James Yoo More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...