Morphological features of IFN-γ-stimulated mesenchymal stromal cells predict overall immunosuppressive capacity

Proc Natl Acad Sci U S A. 2017 Mar 28;114(13):E2598-E2607. doi: 10.1073/pnas.1617933114. Epub 2017 Mar 10.


Human mesenchymal stromal cell (MSC) lines can vary significantly in their functional characteristics, and the effectiveness of MSC-based therapeutics may be realized by finding predictive features associated with MSC function. To identify features associated with immunosuppressive capacity in MSCs, we developed a robust in vitro assay that uses principal-component analysis to integrate multidimensional flow cytometry data into a single measurement of MSC-mediated inhibition of T-cell activation. We used this assay to correlate single-cell morphological data with overall immunosuppressive capacity in a cohort of MSC lines derived from different donors and manufacturing conditions. MSC morphology after IFN-γ stimulation significantly correlated with immunosuppressive capacity and accurately predicted the immunosuppressive capacity of MSC lines in a validation cohort. IFN-γ enhanced the immunosuppressive capacity of all MSC lines, and morphology predicted the magnitude of IFN-γ-enhanced immunosuppressive activity. Together, these data identify MSC morphology as a predictive feature of MSC immunosuppressive function.

Keywords: high-content imaging; immunosuppression; interferon-gamma; mesenchymal stromal cells; morphology.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Flow Cytometry
  • Humans
  • Immunosuppression*
  • In Vitro Techniques
  • Interferon-gamma / pharmacology*
  • Mesenchymal Stem Cells / drug effects
  • Mesenchymal Stem Cells / immunology*
  • Principal Component Analysis


  • Interferon-gamma