Label-free biomarkers of human embryonic stem cell differentiation to hepatocytes

Dimitrios Tsikritsis, Alistair Elfick, Andrew Downes, Vlastimil Srsen, Hu Shi, Yuan Wang, Srinivas Velugotla

Research output: Contribution to journalArticlepeer-review

Abstract

Four different label-free, minimally-invasive, live single cell analysis techniques were applied in a quantitative comparison, to characterize embryonic stem cells and the hepatocytes into which they were differentiated. Atomic Force Microscopy measures the cell’s mechanical properties, Raman spectroscopy measures its chemical properties, and dielectrophoresis measures the membrane’s capacitance. We were able to assign cell type of individual cells with accuracies of 91% (Atomic Force Microscopy), 95.5% (Raman spectroscopy), and 72% (Dielectrophoresis). In addition, Stimulated Raman Scattering (SRS) microscopy was able to easily identify hepatocytes in images by the presence of lipid droplets. These techniques, used either independently or in combination, offer label-free methods to study individual living cells. Although these minimally-invasive biomarkers can be applied to sense phenotypical or environmental changes to cells, these techniques have most potential in human stem cell therapies where the use of traditional biomarkers is best avoided. Destructive assays consume valuable stem cells and do not characterize the cells which go on to be used in therapies; whereas immuno-labelling risks altering cell behaviour. We suggest how these four minimally invasive methods can be applied to cell culture, and how they could in future be combined into one microfluidic chip for cell sorting.
Original languageEnglish
Pages (from-to)575–584
JournalCytometry Part A
Volume89
Issue number6
Early online date23 May 2016
DOIs
Publication statusPublished - 2016

Keywords

  • Raman Spectroscopy
  • Stem cell
  • Hepatocyte
  • Label free detection

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