title: Three-dimensional culture alters primary cardiac cell phenotype.
abstract: The directed formation of complex three-dimensional (3D) tissue architecture is a fundamental goal in tissue engineering and regenerative medicine. The growth of cells in 3D structures is expected to influence cellular phenotype and function, especially relative cell distribution, expression profiles, and responsiveness to exogenous signals; however, relatively few studies have been carried out to examine the effects of 3D reaggregation on cells from critical target organs, like the heart. Accordingly, we cultured primary cardiac ventricular cells in a 3D model system using a serum-free medium to test the hypothesis that expression profiles, multicellular organizational pathways, tissue maturation markers, and responsiveness to hormone stimulation were significantly altered in stable cell populations grown in 3D versus 2D culture. We found that distinct multi-cellular structures formed in 3D in conjunction with changes in mRNA expression profile, up-regulation of endothelial cell migratory pathways, decreases in the expression of fetal genes (Nppa and Ankrd1), and increased sensitivity to tri-iodothyronine stimulation when compared to parallel 2D cultures comprising the same cell populations. These results indicate that the culture of primary cardiac cells in 3D aggregates leads to physiologically relevant alterations in component cell phenotype consistent with cardiac ventricular tissue formation and maturation.
authors.affiliation: Nemours Biomedical Research Center, AI duPont Hospital for Children, Wilmington, Delaware 19803, USA. rakins@nemours.org
authors.forename: Robert E
authors.initials: RE
authors.lastname: Akins
authors.forename: Danielle
authors.initials: D
authors.lastname: Rockwood
authors.forename: Karyn G
authors.initials: KG
authors.lastname: Robinson
authors.forename: Daniel
authors.initials: D
authors.lastname: Sandusky
authors.forename: John
authors.initials: J
authors.lastname: Rabolt
authors.forename: Christian
authors.initials: C
authors.lastname: Pizarro
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D015415:Biomarkers / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D002449:Cell Aggregation / Q000187:drug effects / Q000235:genetics; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002465:Cell Movement / Q000187:drug effects / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002478:Cells, Cultured; D016000:Cluster Analysis; D016895:Culture Media, Serum-Free; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D006352:Heart Ventricles / Q000166:cytology; D009132:Muscles / Q000166:cytology; D009206:Myocardium / Q000166:cytology*; D010641:Phenotype; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014284:Triiodothyronine / Q000494:pharmacology
chemical_list: D015415:Biomarkers; D016895:Culture Media, Serum-Free; D014284:Triiodothyronine