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@article{Tolmachov:2005,
author = {Tolmachov, O and Ma, YL and Themis, M and Patel, P and Spohr, H and Macleod, KT and Ullrich, ND and Kienast, Y and Coutelle, C and Peters, NS},
journal = {BMC Cardiovascular Disorders},
title = {Overexpression of connexin 43 using a retroviral vector improves electrical coupling of skeletal myoblasts with cardiac myocytes in vitro.},
url = {http://www.ncbi.nlm.nih.gov/pubmed/16756651},
volume = {25},
year = {2005}
}

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TY  - JOUR
AB  - BACKGROUND:Organ transplantation is presently often the only available option to repair a damaged heart. As heart donors are scarce, engineering of cardiac grafts from autologous skeletal myoblasts is a promising novel therapeutic strategy. The functionality of skeletal muscle cells in the heart milieu is, however, limited because of their inability to integrate electrically and mechanically into the myocardium. Therefore, in pursuit of improved cardiac integration of skeletal muscle grafts we sought to modify primary skeletal myoblasts by overexpression of the main gap-junctional protein connexin 43 and to study electrical coupling of connexin 43 overexpressing myoblasts to cardiac myocytes in vitro.METHODS:To create an efficient means for overexpression of connexin 43 in skeletal myoblasts we constructed a bicistronic retroviral vector MLV-CX43-EGFP expressing the human connexin 43 cDNA and the marker EGFP gene. This vector was employed to transduce primary rat skeletal myoblasts in optimised conditions involving a concomitant use of the retrovirus immobilising protein RetroNectin and the polycation transduction enhancer Transfectam. The EGFP-positive transduced cells were then enriched by flow cytometry.RESULTS:More than four-fold overexpression of connexin 43 in the transduced skeletal myoblasts, compared with non-transduced cells, was shown by Western blotting. Functionality of the overexpressed connexin 43 was demonstrated by microinjection of a fluorescent dye showing enhanced gap-junctional intercellular transfer in connexin 43 transduced myoblasts compared with transfer in non-transduced myoblasts. Rat cardiac myocytes were cultured in multielectrode array culture dishes together with connexin 43/EGFP transduced skeletal myoblasts, control non-transduced skeletal myoblasts or alone. Extracellular field action potential activation rates in the co-cultures of connexin 43 transduced skeletal myoblasts with cardiac myocytes were significantly higher than in the
AU  - Tolmachov,O
AU  - Ma,YL
AU  - Themis,M
AU  - Patel,P
AU  - Spohr,H
AU  - Macleod,KT
AU  - Ullrich,ND
AU  - Kienast,Y
AU  - Coutelle,C
AU  - Peters,NS
PY  - 2005///
TI  - Overexpression of connexin 43 using a retroviral vector improves electrical coupling of skeletal myoblasts with cardiac myocytes in vitro.
T2  - BMC Cardiovascular Disorders
UR  - http://www.ncbi.nlm.nih.gov/pubmed/16756651
VL  - 25
ER  -

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