Scharenberg CW, Harkey MA, Torok-Storb B. The ABCG2 transporter is an efficient Hoechst 33342 efflux pump and is preferentially expressed by immature human hematopoietic progenitors. The ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype. We suggest that initial experiments be performed using this marrow exactly as we describe in order to establish the procedure in your laboratory and to definitively identify the side population (SP) on the flow cytometer. We highlight the value of multicolor phenotyping, the impact of DNA ploidy, and the importance of distinguishing graft versus host cells for an appropriate SP discrimination. These Bis-benzimides were originally developed by Hoechst AG, which numbered all their compounds so that the dye Hoechst 33342 is the 33,342nd compound made by the company. Camargo FD, Chambers SM, Drew E, McNagny KM, Goodell MA. Goodell MA, Brose K, Paradis G, Conner AS, Mulligan RC. Dye efflux studies suggest that hematopoietic stem cells expressing low or undetectable levels of CD34 antigen exist in multiple species. Irene Sales-Pardo, Ariadna Avendaño, Jordi Barquinero, Joan Carles Domingo, Pedro Marin, Jordi Petriz; The Hoechst low-fluorescent profile of the side population: clonogenicity versus dye retention. By continuing you agree to the Copyright © 2020 Elsevier B.V. or its licensors or contributors. 1 The authors used the Hoechst ABCG2-based efflux assay to isolate primitive stem cells from murine bone marrow (BM). Camargo FD, Chambers SM, Drew E, McNagny KM, Goodell MA. Hoechst is a DNA‐binding dye. Search for other works by this author on: Hoechst stains are part of a family of blue fluorescent dyes used to stain DNA. Isolation and functional properties of murine hematopoietic stem cells that are repopulating in vivo. We have read with great interest the recent paper published in H33342 retention experiments use living cells, allowing the investigators to study an active functional process based on the differential efflux of Reduced accumulation of several fluorescent dyes (ie, rhodamine 123) has also been observed in stem cells, and increased dye efflux has been associated with long-term repopulating ability. Copyright © 2006 by The American Society of Hematology
They demonstrate that side population (SP) cells with a Hoechst 33342 low-fluorescent profile (SP low fraction) have a higher clonogenic potential than the rest of the SP, … The evolving concept of a stem cell: entity or function? Hematopoietic stem cells do not engraft with absolute efficiencies. Stem cells have been isolated by their ability to efflux Hoechst 33342 dye and are referred to as the “side population” (SP). Goodell MA, Brose K, Paradis G, Conner AS, Mulligan RC. This protocol describes the application of Hoechst 33342 (bisbenzimide, H 33342; Ho342) staining to identify and purify side-population (SP) stem cells from various human and murine tissue sources (Goodell et al., 1996, 1997; Storms et al., 2000; Jackson et al., 2001). Sca-1(pos) cells in the mouse mammary gland represent an enriched progenitor cell population. We discuss the SP assay and its applications in stem cell biology, with an emphasis on the technical challenges related to sample preparation, data acquisition, analysis, and interpretation. The side population (SP) assay has been utilized as a method for isolation and characterization of normal and cancer stem cells from a variety of tissues.


In addition to its use in fluorescence microscopy and image analysis, Hoechst 33342 is commonly used for flow cytometric applications, such as cell cycle analysis and stem cell side population identification. Isolation and functional properties of murine hematopoietic stem cells that are replicating in vivo. Keywords: Cancer stem cells, Side population, Fluorescent probes, ABC transporters, JC-1, Hoechst 33342 Background Metabolic alteration in cancer cells has been considered to be linked to cancer progression and drug resistance, and thus characterized intensively for a long time as a potential therapeutic target [ 1 ].

ScienceDirect ® is a registered trademark of Elsevier B.V.Critical Appraisal of the Side Population Assay in Stem Cell and Cancer Stem Cell ResearchCopyright © 2011 Elsevier Inc. All rights reserved.ScienceDirect ® is a registered trademark of Elsevier B.V. The “Side Population” (SP) discrimination assay is a flow cytometry method used to detect stem cells based on the dye efflux properties of ABC transporters. Uchida N, Fujisaki T, Eaves AC, et al. Zhou S, Schuetz JD, Bunting KD, et al. Search for other works by this author on: Search for other works by this author on: The Hoechst side population (SP) technique is an important methodology for identifying stem cells and early progenitors in rodent, nonhuman primate, and human hematopoietic and nonhematopoietic tissues [1, 2, 3 – 4].In this method, the cell‐permeable DNA‐binding dye Hoechst 33342 is passively loaded into cells. Fried J, Doblin J, Takamoto S, Perez A, Hansen H, Clarkson B. Thus, the highest levels of multidrug transporters can be found in CD34For the SP assay, laser power and alignment, filter settings, sample collection, cell viability, We have described a method for isolating hematopoietic stem cells (HSCs) from murine bone marrow utilizing a Hoechst 33342 (Data on the toxicity of H33342 have been available for decades, as it is well known that H33342 is powerful inhibitor of DNA sythesis.Even though Hoechst staining does involve a great deal of viability loss in nonhematopoietic tissues, Hematopoietic stem cells do not engraft with absolute efficiencies. the Hoechst 33342 dye is specific for DNA binding, ribonuclease treatment is not needed to avoid nonspecific RNA staining. Effects of Hoechst 33342 on survival and growth of two tumor cell lines and on hematopoietically normal bone marrow cells. Search for other works by this author on: Van Zant G, Fry CG.


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