66 Fabrication of Engineered Pulsatile 3-Dimensional Scaffold-Free Cardiac Tissue from Embryonic Stem Cells

Abstract

Purpose Transplantation of healthy regenerative cardiomyocytes into the injured heart is effective option to improve cardiac function. The optimal transplantation of cardiomyocytes must be developed. The aim of this study is to fabricate scaffold-free 3-Dimensional pulsatile cardiac tissue by nobel tissue engineering technology. Methods and Materials Approximately two thousands of mouse embryonic stem cells (ESc) derived cardiomyocyte were formed into spheroids. To construct 3-dimensional cardiac tissue, a large number of spheroids are fused via our nobel technology, named bio-rapid prototyping system (BRP). This technolog is based on the original characteristic of cells to form spheroid cell aggregates. The constructs were produced semi-automatically by stabbing spheroids with numerous needles. Finally, we succeeded in fabricating scaffold-free contractile cardiac tissue 3×1×1mm in diameter. Electrophysiological and histological analysis are performed to evaluate the function of the constructs. Results Electrophysiological studies revealed that electrical spikes detected on the construct. Immunofluorescent images with anti-connexin43 antibody demonstrated that gap junctions also established within the construct within 24 hours. This constructs continued pulsation for two weeks in vitro. Figure options Download full-size image Download high-quality image (99 K) Download as PowerPoint slide Conclusions These results demonstrate that electrically communicative pulsatile 3-D cardiac constructs were achieved in vitro by fusing cardiomyocytes spheroids. We concluded that cardiac tissue engineering based on this technology may prove useful for heart model fabrication and cardiovascular tissue repair.