Abstract
This article describes fabrication of a customizable bioreactor, which comprises a perfusion system and coverslip-based tissue culture chamber that allow centimeter-scale vascularized or otherwise canalized tissue constructs to be maintained in weeks long static and/or perfusion culture at an exceptionally low cost, with intermittent live imaging and media sampling capabilities. The perfusion system includes a reusable polydimethylsiloxane (PDMS) lid generated from a three-dimensional (3D)-printed poly-lactic acid (PLA) mold and several lengths of perfusion tubing. The coverslip tissue culture chamber includes PDMS components built with 3D-printed PLA molds, as well as 3D-printed PLA frames and glass coverslips that house perfusable hydrogel constructs. As proof of concept, we fabricated a vascularized hydrogel construct, which was subjected to static and perfusion tissue culture, as well as flow studies using fluorescent beads and widefield fluorescent microscopy. This system can be readily reproduced, promoting the advancement of tissue engineering and regenerative medicine research.
Impact statement
This bespoke bioreactor system allows weeks long (and potentially longer) culture of vascularized or otherwise canalized centimeter-scale cell containing hydrogel constructs, while also permitting live imaging during perfusion cell culture, with intermittent sampling of media for analysis. Furthermore, this system is low-cost, relying on three-dimensional-printed components using poly-lactic acid filament, and molding polydimethylsiloxane to form autoclavable chambers and lids. Using the methods and printing code described herein, other laboratories may readily replicate this system to advance their perfusion-based tissue engineering projects.
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