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24-h Langendorff-perfused neonatal rat heart used to study the impact of adenoviral gene transfer
1 Institute of Experimental and Clinical Pharmacology, University Erlangen-Nürnberg, 91054 Erlangen, Germany; and 2 Institute of Experimental and Clinical Pharmacology, University Hospital Hamburg-Eppendorf, 20246 Hamburg, Germany Submitted 5 November 2002 ; accepted in final form 27 February 2003 T...
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Published in: | American journal of physiology. Heart and circulatory physiology 2003-08, Vol.285 (2), p.H907-H914 |
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Main Authors: | , , , , |
Format: | Article |
Language: | English |
Subjects: | |
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Online Access: | Get full text |
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Summary: | 1 Institute of Experimental and Clinical
Pharmacology, University Erlangen-Nürnberg, 91054 Erlangen, Germany; and
2 Institute 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
-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 135155 beats/min and developed active
force of 12 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 711 mmHg. Perfusion with a LacZ-encoding
adenovirus at 3 x 10 8 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 ). |
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ISSN: | 0363-6135 1522-1539 |
DOI: | 10.1152/ajpheart.00856.2002 |