Three-dimensional engineered heart tissue from neonatal rat cardiac myocytes

Biotechnol Bioeng. 2000 Apr 5;68(1):106-14.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae
  • Animals
  • Animals, Newborn
  • Calcium / pharmacology
  • Cardiotonic Agents / pharmacology
  • Gene Expression
  • Gene Transfer Techniques
  • Genes, Reporter
  • Isoproterenol / pharmacology
  • Muscle Fibers, Skeletal / cytology*
  • Muscle Fibers, Skeletal / drug effects
  • Myocardial Contraction / drug effects
  • Myocardium / cytology*
  • Organ Culture Techniques / instrumentation
  • Organ Culture Techniques / methods*
  • Rats
  • beta-Galactosidase / genetics

Substances

  • Cardiotonic Agents
  • beta-Galactosidase
  • Isoproterenol
  • Calcium