RRC ID 51836
著者 Suga M, Tachikawa S, Tateyama D, Ohnuma K, Furue MK.
タイトル Imaging-cytometry revealed spatial heterogeneities of marker expression in undifferentiated human pluripotent stem cells.
ジャーナル In Vitro Cell Dev Biol Anim
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.
巻・号 53(1)
ページ 83-91
公開日 2017-1-1
DOI 10.1007/s11626-016-0084-3
PII 10.1007/s11626-016-0084-3
PMID 27573412
PMC PMC5258813
MeSH Biomarkers / metabolism* Cell Differentiation* Cell Shape Colony-Forming Units Assay Flow Cytometry / methods* Gene Expression Profiling Gene Expression Regulation Humans Induced Pluripotent Stem Cells / cytology* Induced Pluripotent Stem Cells / metabolism*
IF 1.665
引用数 2
リソース情報
ヒト・動物細胞 201B7(HPS0063) 253G1(HPS0002)