Limited plasticity of mesenchymal stem cells cocultured with adult cardiomyocytes

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...

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Bibliographic Details
Published in:Journal of cellular biochemistry 2007-01, Vol.100 (1), p.86-99
Main Authors: Gallo, Maria Pia, Ramella, Roberta, Alloatti, Giuseppe, Penna, Claudia, Pagliaro, Pasquale, Marcantoni, Andrea, Bonafé, Francesca, Losano, Gianni, Levi, Renzo
Format: Article
Language:English
Subjects:
Actinin - metabolism
Animals
Animals, Genetically Modified
Boron Compounds
Calcium - metabolism
Calcium Channel Blockers - pharmacology
calcium channels
Calcium Channels - physiology
cardiomyocytes
Cell Differentiation
Cells, Cultured
Coculture Techniques
Connexin 43 - metabolism
Dihydropyridines
Fluorescent Dyes
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
Ion Channel Gating
mesenchymal stem cells
Mesenchymal Stromal Cells - cytology
Mesenchymal Stromal Cells - physiology
Myocytes, Cardiac - cytology
Myocytes, Cardiac - physiology
Myosins - metabolism
Nifedipine - pharmacology
Rats
Sarcomeres - metabolism
sarcomeric proteins
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Summary: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. J. Cell. Biochem. 100: 86–99, 2007. © 2006 Wiley‐Liss, Inc.
ISSN:0730-2312
1097-4644
DOI:10.1002/jcb.21012