Intermittent Fasting Extends Life of Liver Cells

Primary human liver cells (PHHs) are crucial for testing harmful drug reactions in human patients. Normally, these cells lose their functions quickly in simple cultures, but when grown with other supportive cells, they last 2-4 weeks. However, since liver cells in the body function for much longer (200-400 days), we want to extend their functional life in the lab to study long-term drug effects and diseases.


Fasting has been shown to improve the health & longevity of tissues like the liver. We thought that mimicking fasting in cell cultures might activate beneficial pathways and extend the life of PHHs. To do this, we periodically removed serum and hormones from a 'micropatterned coculture' of liver cells, which is a special arrangement of liver cells surrounded by connective tissue cells (fibroblasts) for support. A weekly 2-day starvation period extended the life of PHHs to over 6 weeks, compared to just 3 weeks without fasting. This fasting also improved the function of simpler 'monoculture' liver cells for 2 weeks.


In the 'micropatterned cocultures', fasting activated a key energy-regulating protein called AMPK and prevented the supportive fibroblast cells from overgrowing, helping maintain liver cell structure (polarity). Similar benefits were seen when we activated AMPK with a drug called metformin or stopped the growth of supportive cells with another drug, mitomycin-C.


Finally, liver cells in the fasting cultures were better at predicting drug toxicity and had higher drug-processing activities even after 5 weeks. In summary, intermittent fasting in cell cultures extends the functional life of liver cells, making them more useful for drug testing.


Publication >> Intermittent Starvation Extends the Functional Lifetime of Primary Human Hepatocyte Cultures

October 18, 2025
𝗦𝗶𝗺𝗽𝗹𝗲 𝗦𝘂𝗺𝗺𝗮𝗿𝘆: This study explores how we can improve lab-grown liver cells for medical research and drug testing. The MTMLab team works with induced pluripotent stem cells (iPSCs) - special cells that can be transformed into liver-like cells - because real human liver cells are hard to obtain. However, these lab-grown liver cells don't function as well as mature adult liver cells. The research team discovered that the surface environment where these cells grow is crucial for their development. We created tiny fiber scaffolds made from different materials like collagen, decellularized livers, and chitosan that mimic the natural structure around liver cells. When liver cells were grown on these specially designed nanofibers for three weeks, they displayed higher function compared to cells grown on standard surfaces. Our key finding was that both the material composition and the nanoscale fiber structure were important - stiffer synthetic fibers or softer materials without the appropriate topography or composition prevented proper cell maturation. This research helps create better lab models of human liver tissue that can be used for testing new drugs and studying liver diseases more effectively.

2025 BMES Annual Meeting - MTMLab Member Presentations!

October 7, 2025
Owen Lally Modeling the synergistic effects of alcohol and fats on liver disease via engineered cocultures In Vitro Liver Toxicology Testing of Rat and Dog Hepatocytes to Reduce In Vivo Regulatory Requirements Nathan Shelton Enhancing the Functions and Hepatitis B Virus Infectability of Primary Human Hepatocytes Protein Microarrays to Probe Synergistic Effects of Extracellular Matrix Composition and Stiffness on Liver Macrophages Lesly Villarreal Engineering a 3D Placental Trophoblast Invasion Platform Via Droplet Microfluidics Gas-permeable Plates to Model Synergetic Effects of Oxygen and Endothelial Factors on Liver Zonation Emanuele Spanghero Modeling the Interplay Between Liver and Heart Diseases via a Human Dual-Organ Platform Engineering High Cell Density Beating Cords of Cardiomyocytes and Fibroblasts via Photopatterned Alginate

MTMLab Research Summary: Improving Lab-Grown Liver Cell Maturation

May 7, 2025
Our latest study addresses a critical challenge in liver tissue engineering: stem cell-derived liver cells (iHeps) typically remain functionally immature, limiting their usefulness for drug testing and disease modeling. Our research team created 3D microtissues using droplet microfluidics technology by: • Encapsulating iHeps in tiny collagen gel droplets (~250 μm diameter) • Coating these structures with various non-parenchymal cells (NPCs) • Testing different combinations and sequences of supporting cells Key findings: 1) Embryonic fibroblasts and liver sinusoidal endothelial cells (LSECs) produced the most mature iHeps compared to other tested cell types 2) Sequential application of cell signals (embryonic fibroblasts first, then LSECs) yielded optimal maturation 3) Specific growth factors like stromal-derived factor-1 alpha were identified as important maturation enhancers 4) Gene expression analysis confirmed that LSEC/iHep microtissues closely resembled adult human liver cells This platform enables researchers to identify critical cellular interactions and molecular signals that drive liver cell maturation, providing valuable insights for developing more physiologically relevant liver models for drug screening and regenerative medicine applications. https://www.sciencedirect.com/science/article/pii/S174270612500193X SIMPLE SUMMARY: Embryonic fibroblasts and liver sinusoidal endothelial cells dramatically improved iHep maturation compared to other cell types tested, producing more functionally mature liver cells. Sequential application proved crucial—adding embryonic fibroblasts first, followed by endothelial cells, yielded optimal maturation. Specific growth factors including stromal-derived factor-1 enhanced this process. This research enables creation of more authentic mini-liver tissues that function like human liver. These improved models support better drug testing, disease research, and regenerative medicine applications.