Perfusion-based microfluidic device for three-dimensional dynamic primary human hepatocyte cell culture in the absence of biological or synthetic matrices or coagulants

Abstract

We describe a perfusion-based microfluidic device for three-dimensional (3D) dynamic primary human hepatocyte cell culture. The microfluidic device was used to promote and maintain 3D tissue-like cellular morphology and cell-specific functionality of primary human hepatocytes by restoring membrane polarity and hepatocyte transport function in vitro without the addition of biological or synthetic matrices or coagulants. A unique feature of our dynamic cell culture device is the creation of a microenvironment, without the addition of biological or synthetic matrices or coagulants, that promotes the 3D organization of hepatocytes into cord-like structures that exhibit functional membrane polarity as evidenced by the expression of gap junctions and the formation of an extended, functionally active, bile canalicular network.

Chip-based liver equivalents for toxicity testing – organotypicalness versus cost-efficient high throughput

“…However, unlike other perfusion-based microdevices, the bottom of the cell culture chamber was patterned with microstructures, which provided an additional control of hepatocyte polarity. 66 In contrast to Toh et al, the channel is 100 mm wide and hosts 10 4 cells, corresponding to 0.1 human liver lobule. Cell culture media could not only be perfused through the cell compartment from the side-channels, but also via these patterned microstructures on the bottom, minimising cell spreading and cell-surface interaction.…”

“…An extended bile canalicular structure and the formation of gap junctions between the 3D structured cells could be shown. 66 A clear segregation of bile within the cord structures could be achieved. The patterned microstructures at the bottom are the first step toward technical approaches allowing the separation of bile.…”

“…2e). 66 Cells were seeded in a microfluidic device similar to the one described. 64,62 A series of retention pillars formed a microchannel centred between two side-channels.…”

Organ-on-a-Chip Platforms for Drug Delivery and Cell Characterization: A Review

“…(54)(55)(56) The use of a perfused multiwell plate enabled the study of drug toxicity and metabolism [ Fig. 4(b)].…”

Exploitation of physical and chemical constraints for three-dimensional microtissue construction in microfluidics

“…This would then ensure the proper development of the intercellular structures such as the bile canaliculi or sinusoids in case of hepatocytes and lacunae and canaliculi for housing the osteocyte body and osteocyte processes, respectively. 193,219 The bottom-up approach in tissue engineering also needs to be coupled with the top-down approach for solving the real life problems in therapeutics. This would be possible either by multiplexing of bottom-up microtissues or by incorporating bottom-up control features in the top-down scaffolds.…”