Limited plasticity of mesenchymal stem cells cocultured with adult cardiomyocytes
| Autor: | Francesca Bonafè, Pasquale Pagliaro, Andrea Marcantoni, Claudia Penna, Giuseppe Alloatti, Gianni Losano, Maria Pia Gallo, Renzo Levi, Roberta Ramella |
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| Přispěvatelé: | Gallo MP, Ramella R, Alloatti G, Penna C, Pagliaro P, Marcantoni A, Bonafé F, Losano G, Levi R |
| Rok vydání: | 2007 |
| Předmět: |
Boron Compounds
Sarcomeres Dihydropyridines Nifedipine Cellular differentiation Green Fluorescent Proteins sarcomeric proteins chemistry.chemical_element cardiomyocytes Myosins Biology Calcium Biochemistry Calcium in biology Animals Genetically Modified mesenchymal stem cells calcium channels cardiomyocytes mesenchymal stem cells calcium channels sarcomeric proteins Animals Myocyte Actinin Myocytes Cardiac Molecular Biology Cells Cultured Fluorescent Dyes Calcium signaling Calcium metabolism Voltage-dependent calcium channel Mesenchymal stem cell Cell Differentiation Cell Biology Calcium Channel Blockers Coculture Techniques Rats Cell biology chemistry Connexin 43 Ion Channel Gating |
| Zdroj: | Journal of Cellular Biochemistry. 100:86-99 |
| ISSN: | 1097-4644 0730-2312 |
| DOI: | 10.1002/jcb.21012 |
| Popis: | In order to assess, in a controlled in vitro model, the differentiation potential of adult bone marrow derived stem cells we have developed a coculture procedure using adult rat cardiomyocytes and mesenchymal stem cells (MSCs) from transgenic GFP positive rats. We investigated in the cocultured MSCs the time course of cellular processes that are difficult to monitor in in vivo experiments. Adult rat cardiomyocytes and adult rat MSCs were cocultured for up to 7 days and analyzed by confocal microscopy. Several markers were studied by immunofluorescence technique. The fluorescent ST-BODIPY-Dihydropyridine was used to label calcium channels in living cells. Intracellular calcium was monitored with the fluorescent probe X-Rhod-1. Immunofluorescence experiments showed the presence of connexin-43 between cardiomyocytes and MSCs and between MSCs, while no sarcomeric structures were observed at any time of the coculture. We looked at the expression of calcium channels and development of voltage-dependent calcium signaling in cocultured MSCs. MSCs showed a time-dependent increase of labeling of ST-BODIPY-Dihydropyridine, reaching a relatively strong level after 72 h of coculture. The treatment with a non-fluorescent DHP, Nifedipine, completely abolished ST-BODIPY labeling. We investigated whether depolarization could modulate intracellular calcium. Depolarization-induced calcium transients increased in MSCs in relation to the coculture time. We conclude that MSCs cocultured with adult cardiomyocytes present preliminary evidence of voltage-dependent calcium modulation uncoupled with the development of nascent or adult myofibrils, thus showing a limited lineage specification and a low plasticity to differentiate in a full cardiomyocyte-like phenotype. |
| Databáze: | OpenAIRE |
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