Abstract 19821: 3D Tri-culture Model Promotes Enhanced Mechanical Forces and Sustained Long-Term Contractility of Human Pluripotent Stem Cell-Derived Cardiomyocytes
BACKGROUND: Engineered myocardial tissue represents a promising methodology for cardiac repair and regeneration, but the survival and contractility of the cardiomyocytes (CMs) in the engineered tissue remains a limiting factor. Since native CMs interact with endothelial and stromal cells, we generated a three-dimensional (3D) engineered myocardial tissue to study the role of cell-cell interactions on CM survival and function.
HYPOTHESIS: Using human pluripotent stem cells (hPSCs) as a therapeutic cell source, we hypothesize that 3D tri-culture of hPSC-derived CMs (hCMs), hPSC-derived endothelial cells (hECs), and human amniotic mesenchymal stem cells (hAMSCs) will enable sustained, long-term contractility of the hCM population.
METHODS AND RESULTS: We engineered myocardial tissues by culturing combinations of the three cell populations, within a 3D hydrogel for up to 6 weeks as follows: 1) hCMs only (mono-culture), 2) hCMs + hECs (co-culture), 3) hCMs + hAMSCs (co-culture), and 4) hCMs + hECs + hAMSCs (tri-culture). Timelapse imaging was used to compare contractility frequency and beating area, troponin T protein expression using immunofluorescence staining, and cardiac gene expression by qPCR. Quantification of contractility demonstrated enhanced function of hCMs in tri-culture. Expression of cTNT was up-regulated in the tri-culture vs. other culture conditions, which was supported by qPCR measurement of cardiac genes related to maintenance of CM structure (TNNT2 or β-myosin heavy chain, P < 0.01). Enhanced hCM behavior in the tri-culture was further supported by 3-5 fold increases in early and mature cardiac genes (NKX2-5, ISL1, MLC2a, MLC2v, P < 0.05) and electrophysiological properties, including gap junctions (Cx45 and Cx43) and voltage dependent ion channels (Cav1.2 and Kir2.1 *P < 0.05) when compared to other culture conditions.
CONCLUSION: These findings demonstrate that the 3D tri-culture system enabled significantly greater long-term contractility, sustained survival, and enhanced cardiac lineage-related gene expression of hCMs when compared to mono- and co-culture conditions. This study highlights the role of 3D hydrogels and cell-cell interactions to maintain cardiac phenotype and survival.
Author Disclosures: K. Nakayama: None. P. Burridge: Research Grant; Modest; 12POST12050254. S. Metzler: None. O. Abilez: None. C. Simmons: None. M. Bruce: None. Y. Matsuura: None. P. Kim: None. J. Wu: None. M. Butte: None. P. Yang: Research Grant; Modest; 5 UM 1HL087318-08, 5R01HL097516-02. N. Huang: Research Grant; Modest; 1249008, W81XWH-12-C-0111.
This research has received full or partial funding support from the American Heart Association.
- © 2014 by American Heart Association, Inc.