AJP - Heart Fuel your research with LabChart
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Am J Physiol Heart Circ Physiol 285: H907-H914, 2003. First published March 27, 2003; doi:10.1152/ajpheart.00856.2002
0363-6135/03 $5.00
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
285/2/H907    most recent
00856.2002v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via ISI Web of Science (6)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wiechert, S.
Right arrow Articles by Eschenhagen, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wiechert, S.
Right arrow Articles by Eschenhagen, T.

INNOVATIVE METHODOLOGY

24-h Langendorff-perfused neonatal rat heart used to study the impact of adenoviral gene transfer

S. Wiechert,1 A. El-Armouche,2 T. Rau,2 W. -H. Zimmermann,2 and T. Eschenhagen2

1Institute of Experimental and Clinical Pharmacology, University Erlangen-Nürnberg, 91054 Erlangen, Germany; and 2Institute of Experimental and Clinical Pharmacology, University Hospital Hamburg-Eppendorf, 20246 Hamburg, Germany

Submitted 5 November 2002 ; accepted in final form 27 February 2003

The human genome project has increased the demand for simple experimental systems that allow the impact of gene manipulations to be studied under controlled ex vivo conditions. We hypothesized that, in contrast to adult hearts, neonatal hearts allow long-term perfusion and efficient gene transfer ex vivo. A Langendorff perfusion system was modified to allow perfusion for >24 h with particular emphasis on uncompromised contractile activity, sterility, online measurement of force of contraction, inotropic response to {beta}-adrenergic stimulation, and efficient gene transfer. The hearts were perfused with serum-free medium (DMEM + medium 199, 4 + 1) supplemented with hydrocortisone, triiodothyronine, ascorbic acid, insulin, pyruvate, L-carnitine, creatine, taurine, L-glutamine, mannitol, and antibiotics recirculating (500 ml/2 hearts) at 1 ml/min. Hearts from 2 day-old rats beat constantly at 135–155 beats/min and developed active force of 1–2 mN. During 24 h of perfusion, twitch tension increased to ~165% of initial values (P < 0.05), whereas the inotropic response to isoprenaline remained constant. A decrease in total protein content of 10% and histological examination indicated moderate edema, but actin and calsequestrin concentration remained unchanged and perfusion pressure remained constant at 7–11 mmHg. Perfusion with a LacZ-encoding adenovirus at 3 x 108 active virus particles yielded homogeneous transfection of ~80% throughout the heart and did not affect heart rate, force of contraction, or response to isoprenaline compared with uninfected controls (n = 7 each). Taken together, the 24-h Langendorff-perfused neonatal rat heart is a relatively simple, inexpensive, and robust new heart model that appears feasible as a test bed for functional genomics.

functional genomics; heart failure; molecular biology


Address for reprint requests and other correspondence: T. Eschenhagen, Martinistr. 52, 20246 Hamburg, Germany (E-mail: t.eschenhagen{at}uke.uni-hamburg.de).






HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Visit Other APS Journals Online
Copyright © 2003 by the American Physiological Society.