On-chip analysis of glycolysis and mitochondrial respiration in human induced pluripotent stem cells

Stefanie Fuchs, Ruben W.J. van Helden, Maury Wiendels, Mees N.S. de Graaf, Valeria V. Orlova, Christine L. Mummery, Berend J. van Meer, Torsten Mayr*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


Recent advances in microfluidic engineering allow the creation of microenvironments in which human cells can be cultured under (patho-)physiological conditions with greater reality than standard plastic tissue culture plates. Microfluidic devices, also called Organs-on-Chip (OoC), allow complex engineering of the cellular compartment, yielding designs in which microfluidic flow can be precisely controlled. However, it is important that cellular physiology is not only controlled but can also be monitored in these devices. Here, we integrated oxygen and pH sensors into microfluidics, allowing close monitoring of the extracellular flux from the cells, enabling constant assessment of features such as glycolysis and mitochondrial oxidative phosphorylation in situ. Using human-induced pluripotent stem cells (hiPSCs) as an exemplar of a highly metabolic and relatively challenging cell type to maintain, we showed that monitoring the extracellular environment allowed rapid optimization of the seeding protocol. Based on the measurements, we implemented earlier and more frequent media refreshment to counteract the rapid acidification and depletion of oxygen. The integrated sensors showed that hiPSCs in the devices exhibited mitochondrial and glycolytic capacity similar to that measured with the Seahorse extracellular flux system, the most widely used standard for these types of assays in conventional cell culture. Under both conditions, hiPSCs showed greater reliance on glycolysis than mitochondrial OXPHOS and the absolute values obtained were similar. These results thus pave the way for the assessment of cell metabolism in situ under conditions of fluidic flow with the same precision and relevance as current standard static cell cultures.

Original languageEnglish
Article number100475
JournalMaterials Today Bio
Publication statusPublished - 15 Dec 2022


  • Extracellular flux
  • Human stem cells
  • Metabolism
  • Organ-on-Chip
  • Oxygen
  • pH
  • Sensors

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomaterials
  • Biomedical Engineering
  • Molecular Biology
  • Cell Biology


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