Establishment of vessel-like structures in long-term three-dimensional tissue culture of myocardium: an electron microscopy study.
To assess whether long-term three-dimensional (3D) tissue culture of myocardium enables the in vitro establishment of vessel-like structures, myocardial tissue from newborn mice was incubated under conditions of 3D culture for at least 3 weeks, and studied by phase-contrast microscopy, conventional histology, immunohistochemistry, and electron microscopy. During 3 weeks of culture, a mean 24.35 +/- 3.74% of all aggregates contracted spontaneously. The contracting aggregates displayed a tissue-like architecture with small basal and apical zones, and a large central zone. The basal and apical zone consisted of immature mesenchymal cells. The underlying shell of the aggregate contained many cardiomyocytes. Vessel-like structures were found concentrated within the aggregates. Immunohistochemistry showed that up to 15% of the cells in the central zone of the aggregate were positive for the endothelial-specific BS-I lectin. Vessel-like structures were formed by cells, which often showed intracytoplasmatic lumena. Surrounding the neocapillaries, structures of a rudimentary basal membrane could be detected. A 3D culture of myocardial tissue permits the establishment of a rudimentary capillary network within the tissue aggregates, which presumably guarantees a sufficient tissue perfusion up to a maximum aggregate diameter of approximately 900 microm.
Watzka
Stefan B C
SB
Division of Cardiothoracic Surgery, Vienna General Hospital, Vienna, Austria. watzka@tin.it
Steiner
Marianne
M
Samorapoompichit
Puchit
P
Gross
Karin
K
Coles
John G
JG
Wolner
Ernst
E
Weigel
Günter
G
D000222:Adaptation, Physiological / Q000502:physiology; D000818:Animals; D000831:Animals, Newborn; D001705:Bioprosthesis; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D006354:Heart, Artificial; D037102:Lectins / Q000378:metabolism; D051379:Mice; D008833:Microcirculation / Q000502:physiology* / Q000648:ultrastructure*; D032383:Myocytes, Cardiac / Q000502:physiology* / Q000648:ultrastructure*; D018919:Neovascularization, Physiologic / Q000502:physiology*; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*
D037102:Lectins