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Introduction
Scale up liver bioreactor was designed for culturing approximately one million cells organized in an array of 3-dimensional microtissue units under constant perfusion conditions. This mass of tissue meets the sensitivity requirements for many important assays that are performed on cells extracted from the scaffold. Analysis of extracted cells at the end of an experiment and samples of cell culture medium during the culture are the primary means for gathering experimental information from the this bioreactor design. Scaled up liver bioreactor can be used, for example, for evaluation of therapeutics including gene therapy vector performance.
Design
The scale up bioreactor has capacity of approximately one million cells. The cells reside in a scaffold with 1000 channels. Each channel is approximately 300 x 300 x 250 microns in size. The main body the bioreactor consists of three compartments. The scaffold is sandwiched between the bottom two compartments. The third compartment functions as an optical window and is removed to seed the cells into the scaffold. Cell seeding is performed by pipetting cell suspension above the scaffold.
Design & Photo: Karel Domansky
Scaffolds
The scaffolds feature a 20 x 50 array of microchannels. The square channels are 300 x 300 x 230 microns while the rounded channels have diamater of 340 microns.
Laser-machined polycarbonate scaffold with a 20 x 50 array of 300 x 300 x 230-micron channels. The two large laser-machined holes on the sides snuggly fit on Dowel pins in the bioreactor body and provide alignment. Photo: Karel Domansky
Tissue perfusion system
The scale up bioreactor is connected by gas-permeable tubing to two external peristaltic pumps that circulate cell culture medium between a reservoir and the bioreactor. The whole unit (bioreactor, reservoir, and pumps) is placed inside a carbon dioxide incubator for temperature and pH control.
Photo: Karel Domansky
3D model
Exploded view showing individual components of the scale up liver bioreactor. Liver tissue residing in the channels of the cell-holding scaffold is perfused by cell culture medium circulated by external pumps.
3D model: Karel Domansky
Fabrication
The three main bodies of the bioreactor were fabricated by CNC machining from polycarbonate. The size of the three parts assembled together is 78 x 54 x 21 mm. The scaffolds were also fabricated from polycarbonate - either by laser machining or by microdrilling. The retaining rings were machined from stainless steel. The gaskets were punched from silicone.
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