title: Quantitative metabolic imaging using endogenous fluorescence to detect stem cell differentiation.
abstract: The non-invasive high-resolution spatial mapping of cell metabolism within tissues could provide substantial advancements in assessing the efficacy of stem cell therapy and understanding tissue development. Here, using two-photon excited fluorescence microscopy, we elucidate the relationships among endogenous cell fluorescence, cell redox state, and the differentiation of human mesenchymal stem cells into adipogenic and osteoblastic lineages. Using liquid chromatography/mass spectrometry and quantitative PCR, we evaluate the sensitivity of an optical redox ratio of FAD/(NADH + FAD) to metabolic changes associated with stem cell differentiation. Furthermore, we probe the underlying physiological mechanisms, which relate a decrease in the redox ratio to the onset of differentiation. Because traditional assessments of stem cells and engineered tissues are destructive, time consuming, and logistically intensive, the development and validation of a non-invasive, label-free approach to defining the spatiotemporal patterns of cell differentiation can offer a powerful tool for rapid, high-content characterization of cell and tissue cultures.
authors.affiliation: Department of Biomedical Engineering, Tufts University 4 Colby Street Medford, MA 02155.
authors.forename: Kyle P
authors.initials: KP
authors.lastname: Quinn
authors.forename: Gautham V
authors.initials: GV
authors.lastname: Sridharan
authors.forename: Rebecca S
authors.initials: RS
authors.lastname: Hayden
authors.forename: David L
authors.initials: DL
authors.lastname: Kaplan
authors.forename: Kyongbum
authors.initials: K
authors.lastname: Lee
authors.forename: Irene
authors.initials: I
authors.lastname: Georgakoudi
mesh_terms: D050156:Adipogenesis; D002454:Cell Differentiation; D005227:Fatty Acids / Q000096:biosynthesis; D006801:Humans; D059630:Mesenchymal Stem Cells; D055432:Metabolomics / Q000379:methods*; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism; D008954:Models, Biological; D057054:Molecular Imaging / Q000379:methods*; D009243:NAD / Q000378:metabolism; D010084:Oxidation-Reduction; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*
chemical_list: D005227:Fatty Acids; D009243:NAD