A microfabricated array bioreactor for perfused 3D liver culture.

Area of Science:

• Biomedical Engineering
• Tissue Engineering
• Cell Biology

Background:

• Developing functional 3D tissue constructs requires advanced bioreactor systems.
• Continuous perfusion and in situ monitoring are crucial for observing tissue development.

Purpose of the Study:

• To design and evaluate a novel bioreactor for 3D tissue morphogenesis and in situ imaging.
• To assess the impact of cell pre-aggregation on tissue development within the bioreactor.

Main Methods:

• Fabrication of silicon-based scaffolds with microchannels using deep reactive ion etching.
• Integration of scaffolds into a perfusion bioreactor with controlled fluid dynamics and shear stress.
• Culture of primary rat hepatocytes, including pre-aggregated spheroids, for up to two weeks.
• In situ monitoring of tissue structure and function using two-photon microscopy.

Main Results:

• The bioreactor successfully supported 3D tissue formation and cell viability under continuous perfusion.
• Cells rearranged into tissue-like structures within the microchannels.
• Pre-aggregation of hepatocytes into spheroids enhanced tissue morphogenesis and cell viability.
• Repeated in situ imaging provided real-time data on tissue development.

Conclusions:

• The developed bioreactor is effective for engineering 3D tissue structures with controlled microenvironments.
• Cell pre-aggregation is a viable strategy to improve tissue development in perfusion bioreactors.
• The system allows for non-invasive, repeated observation of tissue development and function.