Tissue engineering of functional cardiac muscle: molecular, structural, and electrophysiological studies.
The primary aim of this study was to relate molecular and structural properties of in vitro reconstructed cardiac muscle with its electrophysiological function using an in vitro model system based on neonatal rat cardiac myocytes, three-dimensional polymeric scaffolds, and bioreactors. After 1 wk of cultivation, we found that engineered cardiac muscle contained a 120- to 160-microm-thick peripheral region with cardiac myocytes that were electrically connected through gap junctions and sustained macroscopically continuous impulse propagation over a distance of 5 mm. Molecular, structural, and electrophysiological properties were found to be interrelated and depended on specific model system parameters such as the tissue culture substrate, bioreactor, and culture medium. Native tissue and the best experimental group (engineered cardiac muscle cultivated using laminin-coated scaffolds, rotating bioreactors, and low-serum medium) were comparable with respect to the conduction velocity of propagated electrical impulses and spatial distribution of connexin43. Furthermore, the structural and electrophysiological properties of the engineered cardiac muscle, such as cellularity, conduction velocity, maximum signal amplitude, capture rate, and excitation threshold, were significantly improved compared with our previous studies.
Papadaki
M
M
Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge 02139, Massachusetts.
Bursac
N
N
Langer
R
R
Merok
J
J
Vunjak-Novakovic
G
G
Freed
L E
LE
D000818:Animals; D000831:Animals, Newborn; D019149:Bioreactors; D015153:Blotting, Western; D018031:Connexin 43 / Q000096:biosynthesis; D003402:Creatine Kinase / Q000378:metabolism; D052283:Creatine Kinase, MM Form; D003470:Culture Media; D046508:Culture Techniques; D004594:Electrophysiology; D005455:Fluorescent Antibody Technique; D006321:Heart / Q000502:physiology*; D007527:Isoenzymes / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D007797:Laminin; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D009206:Myocardium / Q000166:cytology / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D018031:Connexin 43; D003470:Culture Media; D007527:Isoenzymes; D007797:Laminin; D007770:L-Lactate Dehydrogenase; D003402:Creatine Kinase; D052283:Creatine Kinase, MM Form; D018995:Myosin Heavy Chains