Rapid fusion between mesenchymal stem cells and cardiomyocytes yields electrically active, non-contractile hybrid cells
| Autor: | Nenad Bursac, Woohyun Yoon, Ilya Y. Shadrin, Liqing Li, Neal Shepherd |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
endocrine system
Calcium Channels L-Type Action Potentials Actinin Real-Time Polymerase Chain Reaction Bioinformatics Time-Lapse Imaging Article Cell Fusion Rats Sprague-Dawley Troponin T Downregulation and upregulation Cell Movement Caffeine Animals Humans Myocyte Myocytes Cardiac Cells Cultured Ions Myosin Type II Microscopy Video Multidisciplinary Cell fusion Voltage-dependent calcium channel Chemistry Mesenchymal stem cell Mesenchymal Stem Cells equipment and supplies Coculture Techniques In vitro Rats 3. Good health Cell biology Connexin 43 Calcium Immunostaining |
| Zdroj: | Scientific Reports |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/srep12043 |
| Popis: | Cardiac cell therapies involving bone marrow-derived human mesenchymal stem cells (hMSCs) have shown promising results, although their mechanisms of action are still poorly understood. Here, we investigated direct interactions between hMSCs and cardiomyocytes in vitro. Using a genetic Ca2+ indicator gCaMP3 to efficiently label hMSCs in co-cultures with neonatal rat ventricular myocytes (NRVMs), we determined that 25–40% of hMSCs (from 4 independent donors) acquired periodic Ca2+ transients and cardiac markers through spontaneous fusion with NRVMs. Sharp electrode and voltage-clamp recordings in fused cells showed action potential properties and Ca2+ current amplitudes in between those of non-fused hMSCs and NRVMs. Time-lapse video-microscopy revealed the first direct evidence of active fusion between hMSCs and NRVMs within several hours of co-culture. Application of blebbistatin, nifedipine or verapamil caused complete and reversible inhibition of fusion, suggesting potential roles for actomyosin bridging and Ca2+ channels in the fusion process. Immunostaining for Cx43, Ki67 and sarcomeric α-actinin showed that fused cells remain strongly coupled to surrounding NRVMs, but downregulate sarcomeric structures over time, acquiring a non-proliferative and non-contractile phenotype. Overall, these results describe the phenotype and mechanisms of hybrid cell formation via fusion of hMSCs and cardiomyocytes with potential implications for cardiac cell therapy. |
| Databáze: | OpenAIRE |
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