title: Imaging-cytometry revealed spatial heterogeneities of marker expression in undifferentiated human pluripotent stem cells.
abstract: Human pluripotent stem cells (hPSCs) provide a good model system for studying human development and are expected as a source for both cell-based medical and pharmaceutical research application. However, stable maintenance of undifferentiated hPSCs is yet challenging, and thus routine characterization is required. Flow-cytometry is one of the popular quantitative characterization tools for hPSCs, but it has drawback of spatial information loss of the cells in the culture. Here, we have applied a two-dimensional imaging cytometry that examines undifferentiated state of hPSCs to analyze localization and morphological information of immunopositive cells in the culture. The whole images of cells in a culture vessel were acquired and analyzed by an image analyzer, IN Cell Analyzer 2000, and determined staining intensity of the cells with their positional information. We have compared the expression of five hPSC-markers in four hPSC lines using the two-dimensional imaging cytometry and flow cytometry. The results showed that immunopositive ratios analyzed by the imaging cytometry had good correlation with those by the flow cytometry. Furthermore, the imaging cytometry revealed spatially heterogenic expression of hPSC-markers in undifferentiated hPSCs. Imaging cytometry is capable of reflecting minute aberrance without losing spatial and morphological information of the cells. It would be a powerful, useful, and time-efficient tool for characterizing hPSC colonies.
authors: Laboratory of Stem Cell Cultures, National Institutes of Biomedical Innovation, Health and Nutrition, 7-6-8 Saito-Asagi, Osaka, Ibaraki, 567-0085, Japan.
authors: Mika
authors: M
authors: Suga
authors: Department of Bioengineering, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka, Niigata, 940-2188, Japan.
authors: Saoko
authors: S
authors: Tachikawa
authors: Research & Development Laboratory, NIPRO Corporation, 3023 Nojicho, Kusatsu, Shiga, 525-0055, Japan.
authors: Daiki
authors: D
authors: Tateyama
authors: Department of Bioengineering, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka, Niigata, 940-2188, Japan. kohnuma@vos.nagaokaut.ac.jp.
authors: Kiyoshi
authors: http://orcid.org/0000-0002-2272-3643
authors: K
authors: Ohnuma
authors: Laboratory of Stem Cell Cultures, National Institutes of Biomedical Innovation, Health and Nutrition, 7-6-8 Saito-Asagi, Osaka, Ibaraki, 567-0085, Japan.
authors: Miho K
authors: MK
authors: Furue
mesh_terms: D015415:Biomarkers / Q000378:metabolism*; D002454:Cell Differentiation; D048430:Cell Shape; D003114:Colony-Forming Units Assay; D005434:Flow Cytometry / Q000379:methods*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*
chemical_list: D015415:Biomarkers
keywords: Cytometry; Human pluripotent stem cell; Imaging; Quantitative characterization