Perfusable Apparatus For Thick-tissue Creation And Growth (patch) Of Cardiac Tissue

Abstract

Cardiac tissue engineering has been developed as a potential alternative treatment for heart failure. However, current 3D tissues are limited in size and thickness due to the lack of an effective vascularization method. We have developed a novel bioreactor system to create viable vascularized cardiac tissue from multicellular spheroids using a digital light processing (DLP) 3D bioprinting system. Spheroids were created from induced pluripotent stem cells (iPSC) and cardiac fibroblasts (FB) using special dimple plates for mass production. One centimeter cubic tissues were created from spheroids using a DLP 3D printed mold with vascular channels. The tissue was maintained in a perfusion chamber under regulated flow and pressure following differentiation to cardiac tissue and endothelialization. Mass production of large spheroids (35,000 / tissue, diameter of 395.99 um +/- 101.15 um) was achieved from 170 million iPSCs and 50 million FBs for the creation of 1cm3 cardiac tissue in a 3D printed mold with vascular channels. The cardiac tissues (n=5) were perfused for 20 days under stable pressure of 17.5 +/- 3.05 PSI and flow of 5000 uL/min +/- 1116.42 uL/min. On days 10 and 20, Alamar blue assays showed viability for all five tissues (Alamar blue intensity: Day 10 1.57 +/- 0.15. Day 20 2.21 +/- 0.19). Thick and viable cardiac tissues were created and maintained using a 3D printed vascularized mold and perfusion system for maturation and growth in vitro for 30 days. This technology will open new doors for viable in vitro cardiac tissue creation.