Membrane-based PDMS microbioreactor for perfused 3D primary rat hepatocyte cultures

Area of Science:

• Biomaterials Engineering
• Hepatocyte Cell Culture
• Microfluidic Devices

Background:

• Primary adult rat hepatocytes are crucial for drug metabolism and toxicology studies.
• Current static culture methods do not fully replicate the in vivo liver microenvironment.
• Improving hepatocyte function in vitro is essential for accurate preclinical testing.

Purpose of the Study:

• To develop novel polydimethylsiloxane (PDMS) microbioreactors for enhanced primary adult rat hepatocyte culture.
• To mimic the in vivo liver architecture and blood perfusion using microfluidic systems.
• To evaluate the impact of these microbioreactors on hepatocyte function and viability.

Main Methods:

• Fabrication of PDMS microbioreactors with integrated polyester or custom-made PDMS membranes (5 x 5 µm pores).
• Perfusion culture of primary adult rat hepatocytes within the microbioreactors for 15 days.
• Comparison of hepatocyte function (albumin secretion, ammonium removal) with static cultures in tissue-culture dishes and inserts.

Main Results:

• The microbioreactors successfully supported cell attachment and reorganization over 15 days.
• Albumin secretion increased 7-fold compared to static cultures and 2-fold compared to insert cultures.
• Ammonium removal was enhanced 7-fold in perfused microbioreactors versus static cultures.
• A new technique for fabricating PDMS membranes with precise pore sizes and thinness was established.

Conclusions:

• The developed PDMS microbioreactors effectively mimic in vivo liver architecture and perfusion.
• These advanced culture systems significantly enhance primary hepatocyte function, offering a superior alternative to static cultures.
• The microbioreactors show great promise for applications in drug screening and liver tissue engineering.