Assembly and Priming of the 3D-Printed Bioreactor for Perfusion Culture

Begin assembling the bioreactor under laminar flow by first assembling the two 3D printed easy flow inserts. Then using one insert as the reservoir, and the other as the reaction chamber, proceed to assemble the perfusion system. Attach two connected three-way valves to the reservoir R1 port using a system tube equipped with a one-way valve.

Connect the resulting outlet with the pump head. Then connect the pump head to the reaction chamber's C4 port using similar tubing with a one-way valve. Next, connect the reaction chamber port C1 via soft wall tubing to the resistance channel with a smaller inner diameter.

Create a perfusion return channel by extending the resistance channel with the system tubing, and closing the luminal circulation loop by connecting to the reservoir at R5.Create an overflow channel by connecting the reaction chamber port C3 to the reservoir port R3 using system tubing, Attach a ventilator filter through R2.Construct a pressure dampener by connecting a syringe filled with air and media to the reaction chamber C6.Using a perfusion media, prime the system through the media exchange ports and the reservoir.