Hydrostatic pressure/perfusion culture system designed and validated for engineering tissue

Tissue engineering to replace or repair damaged tissues using three-dimensional cell constructs is a promising approach to promote tissue regeneration de novo. The production of cell constructs is a critical process for maintaining cell viability and phenotypes in vitro prior to surgical treatment. We have developed a novel hydrostatic pressure (HP)/perfusion culture system for three-dimensional cell constructs with application of mechanical stimuli with HP and continuous medium changes. In this study, we tested and validated the performance of this culture system. This systems' performance was stable at a constant HP up to 5 MPa and at a cyclic HP up to 0-5 MPa at 0.5-0.03 Hz. The performance of medium perfusion in the culture chamber showed laminar flow from an inlet into the chamber parallel to the inner walls. Air bubbles on all inner surfaces of the culture chamber caused unstable HP application because air can be compressed with greater ease than water, consequently impacting fluid compression. Air bubbles in a 3D agarose gel model disappeared due to HP over time and the spaces vacated by the air bubbles were replaced with water. Our validated HP/perfusion culture system allows for a well-regulated constant or cyclic HP application in culture medium and can be applied to other 3D tissue culture for engineering tissue.