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Susceptibility of murine induced pluripotent stem cell-derived cardiomyocytes to hypoxia and nutrient deprivation

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Item Type:Article
Title:Susceptibility of murine induced pluripotent stem cell-derived cardiomyocytes to hypoxia and nutrient deprivation
Creators Name:Brodarac, A. and Šarić, T. and Oberwallner, B. and Mahmoodzadeh, S. and Neef, K. and Albrecht, J. and Burkert, K. and Oliverio, M. and Nguemo, F. and Choi, Y.H. and Neiss, W.F. and Morano, I. and Hescheler, J. and Stamm, C.
Abstract:INTRODUCTION: Induced pluripotent stem cell-derived cardiomyocytes (iPS-CMs) may be suitable for myocardial repair. While their functional and structural properties have been extensively investigated, their response to ischemia-like conditions has not yet been clearly defined. METHODS: iPS-CMs were differentiated and enriched from murine iPS cells expressing eGFP and puromycin resistance genes under the control of an alpha-MHC promoter. iPS-CMs maturity and function were characterized by microscopy, rt-PCR, calcium transient recordings, electrophysiology, and mitochondrial function assays, and compared to those from neonatal murine cardiomyocytes (N-CMs). iPS-CMs as well as N-CMs were exposed for 3 h to hypoxia (1% O2) and glucose/serum deprivation (GSD), and viability, apoptosis markers, reactive oxygen species (ROS), mitochondrial membrane potential (Deltapsim) and intracellular stress signaling cascades were investigated. Then, the iPS-CMs response to mesenchymal stromal cell-conditioned medium (MSC-CoM) was determined. RESULTS: iPS-CMs displayed key morphological and functional properties that were comparable to those of N-CMs, but several parameters indicated an earlier iPS-CMs maturation stage. During hypoxia/GSD, iPS-CMs exhibited a significantly higher proportion of poly-caspase-active, 7-AAD- and TUNEL-positive cells than N-CMs. The average mitochondrial membrane potential (Deltapsim) was reduced in "ischemic" iPS-CMs but remained unchanged in N-CMs, ROS production was only increased in "ischemic" iPS-CMs, and oxidoreductase activity in iPS-CMs dropped more rapidly than in N-CMs. In iPS-CMs, hypoxia/GSD led to upregulation of Hsp70 transcripts and decreased STAT3 phosphorylation and total PKCepsilon protein expression. Treatment with MSC-CoM preserved oxidoreductase activity and restored pSTAT3 and PKCepsilon levels. CONCLUSION: iPS-CMs appear to be particularly sensitive to hypoxia and nutrient deprivation. Counteracting the ischemic susceptibility of iPS-CMs with MSC-conditioned medium may help enhance their survival and efficacy in cell-based approaches for myocardial repair.
Keywords:Calcium, Cardiac Myocytes, Cell Differentiation, Cell Hypoxia, Conditioned Culture Media, Electron Microscopy, Fibroblasts, HSP70 Heat-Shock Proteins, Immunohistochemistry, Inbred C57BL Mice, Induced Pluripotent Stem Cells, Mesenchymal Stromal Cells, Mitochondrial Membrane Potential, Phosphorylation, Protein Kinase C-{epsilon}, Reactive Oxygen Species, STAT3 Transcription Factor, Animals, Mice
Source:Stem Cell Research & Therapy
Publisher:BioMed Central
Page Range:83
Date:23 April 2015
Official Publication:https://doi.org/10.1186/s13287-015-0057-6
PubMed:View item in PubMed

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