A novel high-speed production process to create modular components for the bottom-up assembly of large scale tissue engineered constructs

摘要

To replace damaged or diseased tissues, large tissue-engineered constructs can be prepared by assembling modular components in a bottom-up approach. However, a high speed method is needed to produce sufficient numbers of these modules for full-sized tissue substitutes. To this end, we have devised a novel production technique that combines air shearing and a plug flow reactor-style design to rapidly produce large quantities of hydrogel-based (here type I collagen) cylindrical modular components with tunable diameters and length. Using this technique, modules containing NIH 3T3 cells showed greater than 95% viability while endothelial cell surface attachment and confluent monolayer formation was demonstrated. Additionally, the rapidly-produced modules were used to assemble large tissue constructs (> 1 cm³) in vitro. Module building blocks containing luciferase-expressing L929 cells were packed in full size adult rat liver-shaped bioreactors and perfused with cell medium, to demonstrate the capacity to build organ-shaped constructs; bioluminescence demonstrated sustained viability over 3 days. Cardiomyocyte embedded modules were also used to assemble electrically stimulatable contractile tissue.