YIA-01: YIA Session

A PUMP-FREE 3D MICROFLUIDIC PLATFORM FOR LONG TERM DIFFERENTIATION OF HUMAN LIVER PROGENITOR CELLS

Ong, Louis Jun Ye, Chong Lor Huai, Jin Lin, Teng Yao Zhong Edward, Toh Yi-Chin

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

Microfluidic hepatocyte models are attractive for drug testing applications owing to their ability to mimic the liver sinusoid microenvironment and multiplexing capability Most microfluidic hepatocyte culture devices utilized primary human hepatocytes, which are limited by cell number and batch-to-batch variation Human stem cell-derived hepatocytes offer an alternative cell source although the differentiation and maturation of human stem cell derived hepatocytes in a microfluidic platform has not been demonstrated

Here, we report a first instance of differentiating a human liver stem cell source on a microfluidic 3D perfusion culture platform We developed a pump-free 3D microfluidic perfusion device, which contained a micropillar array to immobilize cells threedimensionally in a central cell culture compartment flanked by two side perfusion channels Pump-free constant perfusion flow was achieved by sustained gravity driven flow from two horizontally-orientated media reservoirs, which were positioned at the inlet and outlet of the device at a fixed height difference The polydimethysiloxane (PDMS) microfluidic perfusion device was housed in a support structure fabricated by 3D printing to define the height difference between the inlet and outlet media reservoirs We tailored the device to suit for hepatocyte cultures using computational fluid dynamics simulation to estimate the height difference needed to achieve different perfusion flow rates, which were validated by experimental flow rate measurement using micro-particle image velocimetry To demonstrate that the pump-free 3D microfluidic perfusion device can support the generation of human stem cell derived hepatocytes, we performed on-chip differentiation of HepaRG cells, a human bipotent stem cell that can differentiate into hepatocytes and biliary cells, for up to 19 days. Compared to static 2D differentiated HepaRG-hepatocytes, HepaRGhepatocytes derived under 3D perfusion exhibited higher liver-specific functions. The pump-free 3D microfluidic device potentially offers simple and robust platform to develop highly functional in vitro liver models incorporating human stem cell derived hepatocytes

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