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Next-Gen Organ-on-a-chip technology augments and extends PHH function in vitro


Our partner Lena Biosciences has created a novel approach to a 3D Perfused organ-on-a-chip model. In the following paper, they demonstrate how this model is used to increase CYP450 activity long-term in PHH models for drug metabolism studies.

(*Reference below)

Current challenges in human drug metabolism studies: 

  • Poor activity of drug metabolizing enzymes in vitro remains a bottleneck – therefore faithfully characterizing CYP450 induction and phase 1 metabolism remains a challenge. 
  • PHH, although the gold standard for drug metabolism studies, have issues with viability and CYP450 expression when transferred from in vivo to in vitro.  
  • Oxygen availability is critical for enzyme function – however it is a challenge to increase oxygen without introducing oxidative stress or diluting metabolites. 
  • Current 3D perfused systems have issues with cost, experimental complexity, scalability, and downstream compatibility with assay reagents. 
  • In some systems there is risk of microbial contamination in tubes. 

Lena Biosciences’ proprietary and patented next-generation organ-on-a-chip insert system, PerfusionPal, uses a unique in-well perfusion method that concentrates drug metabolites, and provides both dissolved oxygen and a hemoglobin-like oxygen supply to hepatocytes analogous to the in vivo environment. 


Key Advantages of the PerfusionPal/SeedEZ system 

  • An easy to use, throughput-scalable organ-on-a-chip insert system.  
  • In-well perfusion circumvents the loss of cell signaling and drug metabolites in otherwise one-way flow of perfusate 
  • Mimics oxygen transport by blood using a breathable hemoglobin analog to improve delivery of respiratory gases to cells. 
    • In vivo approx. 98% of oxygen delivery to cells takes place via reversible binding to hemoglobin. 
    • Blood substitute mimics the function of hemoglobin by delivering additional oxygen to parenchymal cells via the perfusing culture medium that models pericellular interstitial flow.  
  • Requires a single tube to operate 48 statistically independent 3D cell culture organ models. 
    • A comparable organ-on-a-chip platform would require 96 tubes, 48 pumps, and 48 media bottles.  




The PerfusionPal is combined with the proprietary SeedEZ scaffolds seeded with cells in medium or hydrogel.  




Key Features of SeedEZ Scaffolds: 

  • The SeedEz is a 500um thick, transparent, hydrophilic, glass micro-fiber scaffold in which cells can be seeded in medium or hydrogel to encourage both cadherin-mediated cell-cell and integrin mediated cell-extracellular matrix signaling. 
  • SeedEz is optimized for long-term growth of 3D cell cultures and complex 3D co-culture models of brain, cancer, liver, bone, connective tissues, etc 
  • SeedEz scaffolds are easy to handle – they can be easily transferred to another dish using sterile forceps without damaging the cultures and taken out of the PerfusionPal insert and transferred to another dish for quantitative assays, cell isolation, and high-resolution imaging. 
  • Compared to hydrogels, SeedEz provides consistent 3D cell cultures with defined dimensions and reproducible cell distribution in x, y, and z 
  • Cultures in SeedEz are also protected because they cannot be aspirated into a pipet tip during media changes as spheroids can, and they cannot be peeled off the dish as hydrogels can in extended culture. 
  • Cells seeded in the scaffold are maintained in statistically independent wells in their own medium that floats atop a high-density blood substitute that is twice as dense as water and in which medium, reagents, and drugs and immiscible and insoluble. 

Feel free to schedule a technical call if you feel the PerfusionPal/SeedEZ system could benefit your research.  

*Shoemaker J.T., Zhang W., Atlas A.I., Bryan R.A., Inman S.W., Vukasinovic J. A 3D Cell Culture Organ-on-a-Chip Platform With a Breathable Hemoglobin Analogue Augments and Extends Primary Human Hepatocyte Functions in vitro. Frontiers in Molecular Biosciences. 2020.
DOI: 10.3389/fmolb.2020.568777