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2018 Abstracts

Line Upon Line: Engineering Strong 3-Dimensional Cardiac Tissue

Joseph Rich, Brigham Young University

The heart is a crucial and complicated organ in our body. Once heart muscles, or Cardiomyocytes (CMs), are damaged, they cannot regenerate. Heart transplants are difficult to come by, and even with current methods of solving heart disease, heart disease is the leading cause of death today in developed countries [1]. One possible solution to the demand of transplants is through tissue engineering patient-specific cardiac tissue. There are significant problems that need to be solved before cardiac tissue can be useful. Two of these problems that we address in our research are the problems of size and vasculature. The purpose of our research is to create strong 3-dimensional samples of cardiac tissue with complete vascularization. Currently we have successfully decellularized porcine cardiac extracellular matrix (ECM), created 300 micrometer thick samples of the ECM, and make beating heart tissue by seeding in CMs derived from human induced pluripotent stem(IPS) cells into the ECM. These heart tissue samples beat for about 3 months. For future research, to increase vascularization and size, we will place CMs derived from IPS cells together with human umbilical vein endothelial cells (HUVECs) and caridofibroblasts (CFs) at a ratio of CMs:HUVECs:CFs, 1:1:5 [2], 1:3:6 [3], and 2:1:1 [3] onto a ECM and stack those ECMs through centrifugation for 15 minutes[4]. The action potential will be measured, and the vasculature will be seen through staining of the tissue using markers CD31 for HUVECs, Troponin T for CMs, α-actinin for CMs’ sarcomeric lengths, and Cx43 connexin 43 for CMs. By developing these cardiac tissue samples size and vasculature, we will get one step closer to creating patient specific heart implants to cure heart disease. [1] Hiroyuki Yamakawa, et. al. “Strategies for Heart Regeneration Approaches Ranging from Induced Pluripotent Stem Cells to Direct Cardiac Reprogramming,” International Heart Journal, vol. 56, no. 1, p. 1-5, 2015. [Online]. Available: https://www.jstage.jst.go.jp/article/ihj/56/1/56_14-344/_article. [24 July 2017]. [2] Rogozhnikov, Dmitry et al. “Scaffold Free Bio-Orthogonal Assembly of 3-Dimensional Cardiac Tissue via Cell Surface Engineering.” Scientific Reports 6 (2016): 39806. PMC. [Online]. Available: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5180231/ [26 Oct. 2017] [3] Polonchuk, Liudmila et al. “Cardiac Spheroids as Promising in Vitro Models to Study the Human Heart Microenvironment.” Scientific Reports 7 (2017): 7005. PMC. [Online]. Available: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5539326/ [26 Oct. 2017] [4] Haraguchi, Yuji et al. “Three-Dimensional Human Cardiac Tissue Engineered by Centrifugation of Stacked Cell Sheets and Cross-Sectional Observation of Its Synchronous Beatings by Optical Coherence Tomography.” BioMed Research International 2017