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Formation of Cell-To-Cell Connection between Bone Marrow Cells and Isolated Rat Cardiomyocytes in a Cocultivation Model
Autoři | |
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Rok publikování | 2014 |
Druh | Článek v odborném periodiku |
Časopis / Zdroj | Journal of Cell Science & Therapy |
Fakulta / Pracoviště MU | |
Citace | |
Doi | http://dx.doi.org/10.4172/2157-7013.1000185 |
Obor | Fyziologie |
Klíčová slova | Bone marrow; Mononuclear cells; Isolated cardiomyocytes; Cocultivation; Connexins; Cell communication |
Popis | Aims: Limited regenerative potential of cardiomyocytes (CMs) causes irreversible changes in heart tissue during pathological processes. However bone marrow mononuclear cells (BM-MNCs) can migrate to this tissue, incorporate to the area of dead or missing myocytes, and improve the global heart function. The mechanism of BM-MSCs’ incorporation and interaction with CMs is not clear. Our aim was to create an in vitro model which would enable to study the interaction of BM-MNCs with CMs and to make a microscopy description of these interactions. Methods and Results: CMs were isolated from adult and newborn rats. BM-MNCs were isolated from bone marrow. BM-MNCs were added to the myocyte culture. Cell-to-cell adherence and Cx43 expression were evaluated by fluorescence microscopy, Ca2+ transients were evaluated in cardiomyocyte-BMC communication under electrical stimulation by fluo-4 fluorescence measurement. Analysis of calcein transport from BM-MNCs to CMs was performed using fluorescence microscopy. Conclusions: The adherence of BM-MNCs to CMs occurred quickly and was stable. Cx43 was detected in contact zones between BM-MNCs and CMs; pairs which displayed Cx43 positivity represented less than 1% from all BM-MNC-cardiomyocyte pairs in the coculture. Conductive structures between CMs and BM-MNCs were formed and verified by imaging calcein transfer and synchronous Ca2+transients. |
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