B1: Regenerative Medicine I


Anik Islam, Ong Louis Jun Ye, Toh Yi-Chin

National University of Singapore, Singapore; Singapore Institute for Neurotechnology, Singapore

Microfluidic 3D perfusion cell cultures are widely employed in developing organ-on-chip applications because they recapitulate tissue microenvironment and are amenable for multiplexed studies However, substantial technical expertise, sophisticated facilities as well as multiple processing steps are required to fabricate, assemble and operate conventional polydimethylsiloxane (PDMS)-based microfluidic perfusion culture device, which often results in the slow adoption of these devices in routine biological studies and industrial applications Additive manufacturing (or 3D printing) has been shown to offer faster and more accessible method for fabricating microfluidic devices for biological application, although there is no report on devices for microfluidic 3D perfusion cell culture In this study, we show a first instance of a portable 3D microfluidic perfusion culture device fabricated using 3D printing techniques Computer aided drawing (CAD) and computation fluid dynamics simulation (CFD) were used to optimize the design of the microfluidic device, which consisted of an orthogonal microfluidic channel network with microstructures to immobilize and culture cells in 3D The device assembly and operation were also designed to be compatible with fluorescence microscopy and independent of external pumps for portability Two modes of 3D printing, PolyJet and stereolithography, were explored for the device fabrication and compared based on the print quality, resolution and the consistency of the printed parts with the actual CAD model and we found that only stereolithography can generate functional prototypes Finally, we performed biological validation by demonstrating 3D perfusion culture of HepG2 (hepatocarcinoma) and patient-derived head and neck tumor spheroids The cell viability in the 3D spheroids were measured by live-dead staining using fluorescence microscopy while liver-specific synthetic functions of HepG2 were measured With this study, a robust design guideline for microfluidic perfusion culture device using 3D printing was established, which would allow for subsequent optimization and customization of microfluidic culture device using 3D printing technology

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