The use of fluorescent indoline dyes for side population analysis
| Autor: | Taichi Shintou, Takeshi Miyazaki, Tomoaki Shirai, Yasuhiko Tabata, Mie Okano, Kohei Watanabe, Tsuyoshi Nomoto, Hiroshi Kohara |
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| Rok vydání: | 2013 |
| Předmět: |
Indoles
Biophysics Bioengineering Biology Flow cytometry Biomaterials Mice Side population Cancer stem cell medicine Animals Cells Cultured Fluorescent Dyes medicine.diagnostic_test Flow Cytometry Hematopoietic Stem Cells Fluorescence Molecular biology Staining Mice Inbred C57BL medicine.anatomical_structure Microscopy Fluorescence Mechanics of Materials Cell culture Ceramics and Composites Benzimidazoles Female Bone marrow Stem cell |
| Zdroj: | Biomaterials. 34:1024-1032 |
| ISSN: | 0142-9612 |
| DOI: | 10.1016/j.biomaterials.2012.10.059 |
| Popis: | Dye efflux assay evaluated by flow cytometry is useful for stem cell studies. The side population (SP) cells, characterized by the capacity to efflux Hoechst 33342 dye, have been shown to be enriched for hematopoietic stem cells (HSCs) in bone marrow. In addition, SP cells are isolated from various tissues and cell lines, and are also potential candidates for cancer stem cells. However, ultra violet (UV) light, which is not common for every flow cytometer, is required to excite Hoechst 33342. Here we showed that a fluorescent indoline dye ZMB793 can be excited by 488-nm laser, equipped in almost all the modern flow cytometers, and ZMB793-excluding cells showed SP phenotype. HSCs were exclusively enriched in the ZMB793-excluding cells, while ZMB793 was localized in cytosol of bone marrow lineage cells. The efflux of ZMB793 dye was mediated by ATP binding cassette (ABC) transporter Abcg2. Moreover, staining properties were affected by the side-chain structure of the dyes. These data indicate that the fluorescent dye ZMB793 could be used for the SP cell analysis. |
| Databáze: | OpenAIRE |
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