207 AN INNOVATIVE APPROACH TO BUILDING A CLINICALLY RELEVANT SIZED TISSUES AND ORGANS FOR RECONSTRUCTION

Abstract

You have accessJournal of UrologyStem Cell Research1 Apr 2012207 AN INNOVATIVE APPROACH TO BUILDING A CLINICALLY RELEVANT SIZED TISSUES AND ORGANS FOR RECONSTRUCTION Jaehyun Kim, Tanner Hill, Sang Jin Lee, James Yoo, and Anthony Atala Jaehyun KimJaehyun Kim Winston Salem, NC More articles by this author , Tanner HillTanner Hill Winston Salem, NC More articles by this author , Sang Jin LeeSang Jin Lee Winston Salem, NC More articles by this author , James YooJames Yoo Winston Salem, NC More articles by this author , and Anthony AtalaAnthony Atala Winston Salem, NC More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.260AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Building a clinically relevant sized tissue or organ using cells requires maintenance of viable cells until host vasculature is established and integrated into the implanted engineered constructs. However, delay in vasculogenesis results in premature cell death due to the inadequate supply of oxygen and nutrients. One potential solution is to develop methods to maintain cell viability over a long-term by downregulating cellular metabolism until host vascularization is established. Adenosine, a nucleoside which functions as an energy transferring molecule, is reported to increase during hypoxia and functions as a modulator of ion-channel arrest. In this study, we attempted to promote cell survival under hypoxic conditions by exploiting the property of adenosine. METHODS Myoblasts (C2C12 cells) designated as the hypoxic group were transferred to the hypoxic chamber (0.1% O2). A group with no adenosine was placed under hypoxia for up to 13 days to demonstrate eventual cell death. Another group receiving daily doses of adenosine (0 to 10 mM) was incubated for up to 7 days under hypoxia and then placed back into normoxic conditions. The metabolic activity of viable cells at each pre-determined time point was assessed using an MTS assay, which measures mitochondrial activity of cells. RESULTS The metabolic activity of cells grown in normoxia increased linearly with respect to time. Hypoxic cells not treated with adenosine showed an increasing metabolic activity for 7 days under hypoxia, but this resulted in eventual cell death. However, when treated with adenosine, cells under hypoxic conditions maintained a steady state of metabolic activity and these cells resumed their normal metabolic activity instantly when normoxic conditions were restored and adenosine was removed at 7 days. As the dose of adenosine increased from 0 to 10 mM, an escalation of steady hypometabolic state was maintained under hypoxic conditions, and the cells resumed their normal metabolic activity after 7 days. CONCLUSIONS We demonstrate the novel concept that cell viability can be maintained by downregulating cellular metabolism under hypoxic conditions. Application of adenosine to cells under hypoxic conditions prolongs survival by decreasing the metabolic activity to a steady hypometabolic state, thus reducing O2 demand. This concept represents a novel method for increasing cellular survival in tissue-engineered constructs during vasculogenesis. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e86 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jaehyun Kim Winston Salem, NC More articles by this author Tanner Hill Winston Salem, NC More articles by this author Sang Jin Lee Winston Salem, NC More articles by this author James Yoo Winston Salem, NC More articles by this author Anthony Atala Winston Salem, NC More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...