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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., Šarić, T., Oberwallner, B., Mahmoodzadeh, S., Neef, K., Albrecht, J., Burkert, K., Oliverio, M., Nguemo, F., Choi, Y.H., Neiss, W.F., Morano, I., 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 |
| ISSN: | 1757-6512 |
| Publisher: | BioMed Central |
| Volume: | 6 |
| Number: | 1 |
| 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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