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Cardiac tissue engineering: Cell seeding, cultivation parameters, and tissue construct characterization
Author(s) -
Carrier Rebecca L.,
Papadaki Maria,
Rupnick Maria,
Schoen Frederick J.,
Bursac Nenad,
Langer Robert,
Freed Lisa E.,
VunjakNovakovic Gordana
Publication year - 1999
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19990905)64:5<580::aid-bit8>3.0.co;2-x
Subject(s) - laboratory flask , tissue engineering , seeding , bioreactor , cell culture , cell , microbiology and biotechnology , biology , biophysics , embryonic stem cell , chemistry , biomedical engineering , biochemistry , botany , medicine , genetics , gene , agronomy
Cardiac tissue engineering has been motivated by the need to create functional tissue equivalents for scientific studies and cardiac tissue repair. We previously demonstrated that contractile cardiac cell–polymer constructs can be cultivated using isolated cells, 3‐dimensional scaffolds, and bioreactors. In the present work, we examined the effects of (1) cell source (neonatal rat or embryonic chick), (2) initial cell seeding density, (3) cell seeding vessel, and (4) tissue culture vessel on the structure and composition of engineered cardiac muscle. Constructs seeded under well‐mixed conditions with rat heart cells at a high initial density ((6–8) × 10 6 cells/polymer scaffold) maintained structural integrity and contained macroscopic contractile areas (approximately 20 mm 2 ). Seeding in rotating vessels (laminar flow) rather than mixed flasks (turbulent flow) resulted in 23% higher seeding efficiency and 20% less cell damage as assessed by medium lactate dehydrogenase levels ( p < 0.05). Advantages of culturing constructs under mixed rather than static conditions included the maintenance of metabolic parameters in physiological ranges, 2–4 times higher construct cellularity ( p ≤ 0.0001), more aerobic cell metabolism, and a more physiological, elongated cell shape. Cultivations in rotating bioreactors, in which flow patterns are laminar and dynamic, yielded constructs with a more active, aerobic metabolism as compared to constructs cultured in mixed or static flasks. After 1–2 weeks of cultivation, tissue constructs expressed cardiac specific proteins and ultrastructural features and had approximately 2–6 times lower cellularity ( p < 0.05) but similar metabolic activity per unit cell when compared to native cardiac tissue. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 64: 580–589, 1999.