Abstract
Exclusion of Hoechst 33342 dye is a characteristic common to stem cells, as well as chemotherapy-resistant cancer cells. Normally, these dye-excluding cells can be sorted from enzymatically dissociated tissues with a UV cell sorter/flow cytometer. UV-flow cytometry can be expensive, time-consuming and not readily available to all laboratories. We have developed a simple, high-throughput 96-well microtiter plate assay by which cell populations can be quickly screened for Hoechst dye uptake and exclusion. The method is compatible with green-fluorescent EGFP expressing cells, often used in stem cell biology. Useful applications for this assay will be the rapid screening of clonal stem cell populations and tumor cells for Hoechst dye uptake.
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References
S. Bhattacharya J.D. Jackson A.V. Das W.B. Thoreson C. Kuszinski J. James S. Joshi I. Ahmad (2003) ArticleTitleDirect identification and enrichment of retinal stem cells/progenitors by Hoechst dye efflux assay Invest. Ophthalmol. Vis. Sci 44 IssueID6 2764–2773 Occurrence Handle12766085
R.D. Cone T. Grodzicker M. Jaramillo (1988) ArticleTitleA retrovirus expressing the 12S adenoviral E1A gene product can immortalize epithelial cells from a broad range of rat tissues Mol. Cell. Biol. 8 1036–1044 Occurrence Handle2966895
L.A. Doyle D.D. Ross (2003) ArticleTitleMultidrug resistance mediated by the breast cancer resistance protein BCRP (ABCG2) Oncogene 22 IssueID47 7340–7358 Occurrence Handle10.1038/sj.onc.1206938 Occurrence Handle14576842
O.S. Frankfurt (1983) ArticleTitleIncreased uptake of vital dye Hoechst 33342 during S phase in synchronized HeLa S3 cells Cytometry 4 IssueID3 216–221 Occurrence Handle10.1002/cyto.990040305 Occurrence Handle6661988
T. Frey (1995) ArticleTitleNucleic acid dyes for detection of apoptosis in live cells Cytometry 21 IssueID3 265–274 Occurrence Handle10.1002/cyto.990210307 Occurrence Handle8582249
M. Goodell K. Brose G. Paradis A.S. Conner R.C. Mulligan (1996) ArticleTitleIsolation and functional properties of murine hematopoetic stem cells that are replicating in vivo J. Exp. Med. 183 1797–1806 Occurrence Handle10.1084/jem.183.4.1797 Occurrence Handle8666936
M.H. Kim J. Turnquist M. Jackson et al. (2002) ArticleTitleThe multidrug resistance transporter ABCG2 (breast cancer resistance protein 1) effluxes Hoechst 33342 and is overexpressed in hematopoietic stem cells Clin. Cancer Res. 8 22–28 Occurrence Handle11801536
S. Lahmy P. Viallet J.M. Salmon (1995) ArticleTitleIs reduced accumulation of Hoechst 33342 in multidrug resistant cells related to P-glycoprotein activity? Cytometry. 19 IssueID2 126–133 Occurrence Handle10.1002/cyto.990190207 Occurrence Handle7743893
M. Maynadie G. Lizard E. Solary P. Roignot P.M. Carli (1994) ArticleTitleMultidrug resistance phenotype evaluation by immunofluorescence and functional tests: comparison of two monoclonal antibodies and three fluorescent dyes in three cells lines Anticancer Res. 14 IssueID6B 2605–2609 Occurrence Handle7872688
G.C. Rice J.W. Gray W.C. Dewey (1985) ArticleTitleFACS analysis of a hyperthermia-induced alteration in Hoechst 33342 permeability and direct measurement of its relationship to cell survival J. Cell Physiol. 122 IssueID3 387–396 Occurrence Handle10.1002/jcp.1041220308 Occurrence Handle3968193
CW Scharenberg M.A. Harkey B. Torok-Storb (2002) ArticleTitleThe ABCG2 transporter is an efficient Hoechst 33342 efflux pump and is preferentially expressed by immature human hematopoietic progenitors Blood 99 507–512 Occurrence Handle10.1182/blood.V99.2.507 Occurrence Handle11781231
G.M. Seigel (1996) ArticleTitleEstablishment of an E1A-immortalized rat retinal cell culture In Vitro Cell Dev. Biol. 32 66–68
G.M. Seigel W. Sun J. Wang D. Hershberger L. Campbell R. Salvi (2004) ArticleTitleNeuronal gene expression and function in the growth-stimulated R28 retinal precursor cell line Curr. Eye Res. 28 257–269 Occurrence Handle10.1076/ceyr.28.4.257.27831 Occurrence Handle15259295
T.W. Sery E.Y. Lee W.H. Lee R. Bookstein V. Wong J.A. Shields J.J. Augsburger L.A.J. Donoso (1990) ArticleTitleCharacteristics of two new retinoblastoma cell lines: WERI-Rb24 and WERI-Rb27 Pediatr. Ophthalmol. Strabismus. 27 IssueID4 212–217
R. Summer D.N. Kotton X. Sun B. Ma K. Fitzsimmons A. Fine (2003) ArticleTitleSide population cells and Bcrp1 expression in lung Am. J. Physiol. Lung Cell. Mol. Physiol. 285 L97–104 Occurrence Handle12626330
B.E. Welm S.B. Tepera T. Venezia T.A. Graubert J.M. Rosen M.A. Goodell (2002) ArticleTitleSca1 positive cells in the mouse mammary gland represent an enriched progenitor cell population Dev. Biol. 245 42–56 Occurrence Handle10.1006/dbio.2002.0625 Occurrence Handle11969254
S. Zhou J.D. Schuetz K.D. Bunting A.M. Colapietro J. Sampath J.J. Morris J. Lagutina G.C. Grosveld M. Osawa H. Nakauchi S.P. Sorrentino (2001) ArticleTitleThe ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype Nat. Med. 7 1028–1034 Occurrence Handle10.1038/nm0901-1028 Occurrence Handle11533706
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Seigel, G.M., Campbell, L.M. High-throughput microtiter assay for Hoechst 33342 dye uptake. Cytotechnology 45, 155–160 (2004). https://doi.org/10.1007/s10616-004-7256-9
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DOI: https://doi.org/10.1007/s10616-004-7256-9