{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T01:03:16Z","timestamp":1790384596147,"version":"4.1.0"},"reference-count":23,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2008,9,4]],"date-time":"2008-09-04T00:00:00Z","timestamp":1220486400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cytometry Pt A"],"published-print":{"date-parts":[[2008,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Side population (SP) analyses and CD133 expression have identified cells with stem\u2010like potential in normal and cancerous tissue. Whether stem\u2010like cells exist in cancer cell lines is hotly debated. We have interrogated the DAOY medulloblastoma cell line with respect to stem\u2010like potential. Vital staining for Hoechst 33342 efflux capacity and CD133 immunophenotyping were performed on DAOY cells to assess the presence of the SP and the CD133 stem cell markers, respectively. SP\/non\u2010SP and CD133<jats:sup>+<\/jats:sup>\/CD133<jats:sup>\u2212<\/jats:sup> DAOY cells were sorted into separate fractions for limiting dilution analysis (tumor sphere assay) and asymmetric division assessment. SP\/non\u2010SP cells were also sorted separately for viability (XTT assay), cell size, cell cycle status, and proliferative capacity (carboxyfluorescein succinimidyl ester (CFSE)) evaluation. A minor proportion of cells displayed either the SP or the CD133<jats:sup>+<\/jats:sup> phenotypes. CD133 expression mapped to both the SP and non\u2010SP compartments, with CD133<jats:sup>+<\/jats:sup> cells being enriched almost fourfold within the non\u2010SP gate. The SP, non\u2010SP, CD133<jats:sup>+<\/jats:sup>, and CD133<jats:sup>\u2212<\/jats:sup> fractions were all capable of reconstituting the original parental DAOY population. Slight clonogenic enrichment was observed in only the SP fraction; however, both CD133<jats:sup>+<\/jats:sup> and CD133<jats:sup>\u2212<\/jats:sup> cells displayed equivalent stem cell\u2010like frequencies. SP cells were resistant to Hoechst 33342\u2010mediated toxicity relative to the parental population and differed from the non\u2010SP cells with respect to increased cell size, decreased S\u2010phase, and slightly decreased proliferative capacity. The multiparametric strategy described in this study revealed that the SP and CD133<jats:sup>+<\/jats:sup> subset may be two independent compartments. Our results highlight the need for new reliable specific cancer stem cell marker(s) as Hoechst 33342 efflux and CD133 expression might not be suitable for selectively isolating cancer stem\u2010like cells from cell lines, as shown for the DAOY cells. As such, care must be used in interpreting therapeutic studies targeting the stem cell compartment of cancer cell lines. \u00a9 2008 International Society for Advancement of Cytometry<\/jats:p>","DOI":"10.1002\/cyto.a.20633","type":"journal-article","created":{"date-parts":[[2008,9,4]],"date-time":"2008-09-04T17:47:34Z","timestamp":1220550454000},"page":"940-948","source":"Crossref","is-referenced-by-count":38,"title":["Flow cytometric characterization of the DAOY medulloblastoma cell line for the cancer stem\u2010like phenotype"],"prefix":"10.1002","volume":"73A","author":[{"given":"Vinit Krishna","family":"Srivastava","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Josephine","family":"Nalbantoglu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2008,9,4]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)81683-9"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/35102167"},{"key":"e_1_2_6_4_2","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-05-3986"},{"key":"e_1_2_6_5_2","first-page":"5821","article-title":"Identification of a cancer stem cell in human brain tumors","volume":"63","author":"Singh SK","year":"2003","journal-title":"Cancer Res"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrc1740"},{"key":"e_1_2_6_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/367645a0"},{"key":"e_1_2_6_8_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.183.4.1797"},{"key":"e_1_2_6_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijrobp.2006.09.037"},{"key":"e_1_2_6_10_2","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-06-0858"},{"key":"e_1_2_6_11_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0307618100"},{"key":"e_1_2_6_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/0012-1606(89)90291-1"},{"key":"e_1_2_6_13_2","doi-asserted-by":"publisher","DOI":"10.1006\/dbio.1998.9192"},{"key":"e_1_2_6_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrc1590"},{"key":"e_1_2_6_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(94)90212-7"},{"key":"e_1_2_6_16_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.192276999"},{"key":"e_1_2_6_17_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.202607599"},{"key":"e_1_2_6_18_2","first-page":"999","article-title":"Multiparameter analysis of transplantable hemopoietic stem cells: I. 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