Pacemaker current gene expression of rat mesenchymal stem cells and identification of mesenchymal stem cells expressing human pacemaker current gene (hHCN2) / 中华心血管病杂志
Chinese Journal of Cardiology
;
(12): 917-921, 2006.
Artículo
en Chino
| WPRIM
| ID: wpr-238491
ABSTRACT
<p><b>OBJECTIVE</b>To study pacemaker current gene expression of mesenchymal stem cells (MSCs) and the electrophysiological property of MSCs expressing human pacemaker current gene.</p><p><b>METHODS</b>Pacemaker current gene expression of MSCs were studied by real-time quantitative polymerase chain reaction (real-time PCR) and pcDNA3-hHCN2 was transfected with Lipofectin 2000 into MSCs. hHCN2 expression at mRNA and at protein levels in the transfected cells were identified by real-time PCR and Western blot, respectively. The ionic currents of cloned hHCN2 (IhHCN2) were recorded and the current characteristics were studied through the whole-cell patch clamp technique.</p><p><b>RESULTS</b>mHCN1, mHCN2, mHCN3, mHCN4 represent (0.08+/-0.01)%, (77.16+/-0.03)%, (0.24+/-0.01)%, (22.53+/-0.02)% of total HCN mRNA in MSCs as determined by real-time PCR. Transfected hHCN2 ionic currents were recorded by whole-cell patch clamp and current density-voltage curves were obtained. The threshold for activation of IhHCN2 was approximately -80 mV and this current could be blocked by Cs+ (4 mmol/L). hHCN2 expression in transfected MSCs was detected both at mRNA and protein levels.</p><p><b>CONCLUSIONS</b>1. mHCN2 and mHCN4 represent the major populations of total HCN mRNA in MSCs. 2. Plasmid pcDNA3-hHCN2 by Lipofectin could be successfully transfected into MSCs with IhHCN2 recorded by whole-cell patch clamp technique, this study provides a basis for future antiarrhythmic gene therapy.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Fisiología
/
Transfección
/
Canales de Potasio
/
Expresión Génica
/
Reacción en Cadena de la Polimerasa
/
Ratas Sprague-Dawley
/
Biología Celular
/
Canales Catiónicos Regulados por Nucleótidos Cíclicos
/
Células Madre Mesenquimatosas
/
Canales Regulados por Nucleótidos Cíclicos Activados por Hiperpolarización
Tipo de estudio:
Estudio pronóstico
Límite:
Animales
/
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Cardiology
Año:
2006
Tipo del documento:
Artículo
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