Organ explant culture of neonatal rat ventricles: a new model to study gene and cell therapy.

Testing cardiac gene and cell therapies in vitro requires a tissue substrate that survives for several days in culture while maintaining its physiological properties. The purpose of this study was to test whether culture of intact cardiac tissue of neonatal rat ventricles (organ explant culture) may...

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Autores principales:	A Dénise den Haan, Marieke W Veldkamp, Diane Bakker, Geert J J Boink, Rob B Janssen, Jacques M T de Bakker, Hanno L Tan
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/83162c2e80884191af240d27694f51ee
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spelling	oai:doaj.org-article:83162c2e80884191af240d27694f51ee2021-11-18T07:53:32ZOrgan explant culture of neonatal rat ventricles: a new model to study gene and cell therapy.1932-620310.1371/journal.pone.0059290https://doaj.org/article/83162c2e80884191af240d27694f51ee2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23516623/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Testing cardiac gene and cell therapies in vitro requires a tissue substrate that survives for several days in culture while maintaining its physiological properties. The purpose of this study was to test whether culture of intact cardiac tissue of neonatal rat ventricles (organ explant culture) may be used as a model to study gene and cell therapy. We compared (immuno) histology and electrophysiology of organ explant cultures to both freshly isolated neonatal rat ventricular tissue and monolayers. (Immuno) histologic studies showed that organ explant cultures retained their fiber orientation, and that expression patterns of α-actinin, connexin-43, and α-smooth muscle actin did not change during culture. Intracellular voltage recordings showed that spontaneous beating was rare in organ explant cultures (20%) and freshly isolated tissue (17%), but common (82%) in monolayers. Accordingly, resting membrane potential was -83.9±4.4 mV in organ explant cultures, -80.5±3.5 mV in freshly isolated tissue, and -60.9±4.3 mV in monolayers. Conduction velocity, measured by optical mapping, was 18.2±1.0 cm/s in organ explant cultures, 18.0±1.2 cm/s in freshly isolated tissue, and 24.3±0.7 cm/s in monolayers. We found no differences in action potential duration (APD) between organ explant cultures and freshly isolated tissue, while APD of monolayers was prolonged (APD at 70% repolarization 88.8±7.8, 79.1±2.9, and 134.0±4.5 ms, respectively). Organ explant cultures and freshly isolated tissue could be paced up to frequencies within the normal range for neonatal rat (CL 150 ms), while monolayers could not. Successful lentiviral (LV) transduction was shown via Egfp gene transfer. Co-culture of organ explant cultures with spontaneously beating cardiomyocytes increased the occurrence of spontaneous beating activity of organ explant cultures to 86%. We conclude that organ explant cultures of neonatal rat ventricle are structurally and electrophysiologically similar to freshly isolated tissue and a suitable new model to study the effects of gene and cell therapy.A Dénise den HaanMarieke W VeldkampDiane BakkerGeert J J BoinkRob B JanssenJacques M T de BakkerHanno L TanPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 3, p e59290 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q A Dénise den Haan Marieke W Veldkamp Diane Bakker Geert J J Boink Rob B Janssen Jacques M T de Bakker Hanno L Tan Organ explant culture of neonatal rat ventricles: a new model to study gene and cell therapy.
description	Testing cardiac gene and cell therapies in vitro requires a tissue substrate that survives for several days in culture while maintaining its physiological properties. The purpose of this study was to test whether culture of intact cardiac tissue of neonatal rat ventricles (organ explant culture) may be used as a model to study gene and cell therapy. We compared (immuno) histology and electrophysiology of organ explant cultures to both freshly isolated neonatal rat ventricular tissue and monolayers. (Immuno) histologic studies showed that organ explant cultures retained their fiber orientation, and that expression patterns of α-actinin, connexin-43, and α-smooth muscle actin did not change during culture. Intracellular voltage recordings showed that spontaneous beating was rare in organ explant cultures (20%) and freshly isolated tissue (17%), but common (82%) in monolayers. Accordingly, resting membrane potential was -83.9±4.4 mV in organ explant cultures, -80.5±3.5 mV in freshly isolated tissue, and -60.9±4.3 mV in monolayers. Conduction velocity, measured by optical mapping, was 18.2±1.0 cm/s in organ explant cultures, 18.0±1.2 cm/s in freshly isolated tissue, and 24.3±0.7 cm/s in monolayers. We found no differences in action potential duration (APD) between organ explant cultures and freshly isolated tissue, while APD of monolayers was prolonged (APD at 70% repolarization 88.8±7.8, 79.1±2.9, and 134.0±4.5 ms, respectively). Organ explant cultures and freshly isolated tissue could be paced up to frequencies within the normal range for neonatal rat (CL 150 ms), while monolayers could not. Successful lentiviral (LV) transduction was shown via Egfp gene transfer. Co-culture of organ explant cultures with spontaneously beating cardiomyocytes increased the occurrence of spontaneous beating activity of organ explant cultures to 86%. We conclude that organ explant cultures of neonatal rat ventricle are structurally and electrophysiologically similar to freshly isolated tissue and a suitable new model to study the effects of gene and cell therapy.
format	article
author	A Dénise den Haan Marieke W Veldkamp Diane Bakker Geert J J Boink Rob B Janssen Jacques M T de Bakker Hanno L Tan
author_facet	A Dénise den Haan Marieke W Veldkamp Diane Bakker Geert J J Boink Rob B Janssen Jacques M T de Bakker Hanno L Tan
author_sort	A Dénise den Haan
title	Organ explant culture of neonatal rat ventricles: a new model to study gene and cell therapy.
title_short	Organ explant culture of neonatal rat ventricles: a new model to study gene and cell therapy.
title_full	Organ explant culture of neonatal rat ventricles: a new model to study gene and cell therapy.
title_fullStr	Organ explant culture of neonatal rat ventricles: a new model to study gene and cell therapy.
title_full_unstemmed	Organ explant culture of neonatal rat ventricles: a new model to study gene and cell therapy.
title_sort	organ explant culture of neonatal rat ventricles: a new model to study gene and cell therapy.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/83162c2e80884191af240d27694f51ee
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Organ explant culture of neonatal rat ventricles: a new model to study gene and cell therapy.

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