title: Morphological features of IFN-γ-stimulated mesenchymal stromal cells predict overall immunosuppressive capacity.
abstract: 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.
authors.affiliation: Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, US Food and Drug Administration, Silver Spring, MD 20993.
authors.forename: Matthew W
authors.initials: MW
authors.lastname: Klinker
authors.affiliation: Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, US Food and Drug Administration, Silver Spring, MD 20993.
authors.forename: Ross A
authors.initials: RA
authors.lastname: Marklein
authors.affiliation: Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, US Food and Drug Administration, Silver Spring, MD 20993.
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Lo Surdo
authors.affiliation: Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, US Food and Drug Administration, Silver Spring, MD 20993.
authors.forename: Cheng-Hong
authors.initials: CH
authors.lastname: Wei
authors.affiliation: Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, US Food and Drug Administration, Silver Spring, MD 20993 steven.bauer@fda.hhs.gov.
authors.forename: Steven R
authors.identifier: 0000-0003-2831-846X
authors.initials: SR
authors.lastname: Bauer
mesh_terms: D005434:Flow Cytometry; D006801:Humans; D007165:Immunosuppression Therapy; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000494:pharmacology*; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000276:immunology*; D025341:Principal Component Analysis
chemical_list: D007371:Interferon-gamma
keywords: high-content imaging; immunosuppression; interferon-gamma; mesenchymal stromal cells; morphology