Three-dimensional engineered heart tissue from neonatal rat cardiac myocytes.
A technique is presented that allows neonatal rat cardiac myocytes to form spontaneously and coherently beating 3-dimensional engineered heart tissue (EHT) in vitro, either as a plane biconcaval matrix anchored at both sides on Velcro-coated silicone tubes or as a ring. Contractile activity was monitored in standard organ baths or continuously in a CO(2) incubator for up to 18 days (=26 days after casting). Long-term measurements showed an increase in force between days 8 and 18 after casting and stable forces thereafter. At day 10, the twitch amplitude (TA) of electrically paced EHTs (average length x width x thickness, 11 x 6 x 0.4 mm) was 0.51 mN at length of maximal force development (L(max)) and a maximally effective calcium concentration. EHTs showed typical features of neonatal rat heart: a positive force-length and a negative force-frequency relation, high sensitivity to calcium (EC(50) 0.24 mM), modest positive inotropic (increase in TA by 46%) and pronounced positive lusitropic effect of isoprenaline (decrease in twitch duration by 21%). Both effects of isoprenaline were sensitive to the muscarinic receptor agonist carbachol in a pertussis toxin-sensitive manner. Adenovirus-mediated gene transfer of beta-galactosidase into EHTs reached 100% efficiency. In summary, EHTs retain many of the physiological characteristics of rat cardiac tissue and allow efficient gene transfer with subsequent force measurement.
Zimmermann
W H
WH
Institut für Experimentelle und Klinische Pharmakologie und Toxikologie, Abteilung für Pharmakologie, Universitäts-Krankenhaus Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany.
Fink
C
C
Kralisch
D
D
Remmers
U
U
Weil
J
J
Eschenhagen
T
T
D000256:Adenoviridae; D000818:Animals; D000831:Animals, Newborn; D002118:Calcium / Q000494:pharmacology; D002316:Cardiotonic Agents / Q000494:pharmacology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D007545:Isoproterenol / Q000494:pharmacology; D018485:Muscle Fibers, Skeletal / Q000166:cytology* / Q000187:drug effects; D009200:Myocardial Contraction / Q000187:drug effects; D009206:Myocardium / Q000166:cytology*; D009924:Organ Culture Techniques / Q000295:instrumentation / Q000379:methods*; D051381:Rats; D001616:beta-Galactosidase / Q000235:genetics
D002316:Cardiotonic Agents; D001616:beta-Galactosidase; D007545:Isoproterenol; D002118:Calcium