The Hoechst low-fluorescent profile of the side population: clonogenicity versus dye retention

Sales-Pardo, Irene; Avendaño, Ariadna; Barquinero, Jordi; Domingo, Joan Carles; Marı́n, Pedro; Pétriz, Jordi

doi:10.1182/blood-2006-03-013615

Cited by 4 publications

(3 citation statements)

References 8 publications

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“…The cell suspension was labeled in the same media with 5 µg/ml of Hoechst 33342 dye (Ho-33342) (Sigma-Aldrich, Spain), either alone or in combination with 100 µM verapamil (Vp) (Sigma-Aldrich, Spain) [26], in a water bath at 37°C for 90–120 minutes. Then, cells were centrifuged for 6 minutes at 4°C, and were resuspended in cold HBSS supplemented with 2%FBS and 10 mM HEPES.…”

Section: Methodsmentioning

confidence: 99%

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Human Endometrial Side Population Cells Exhibit Genotypic, Phenotypic and Functional Features of Somatic Stem Cells

et al. 2010

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During reproductive life, the human endometrium undergoes around 480 cycles of growth, breakdown and regeneration should pregnancy not be achieved. This outstanding regenerative capacity is the basis for women's cycling and its dysfunction may be involved in the etiology of pathological disorders. Therefore, the human endometrial tissue must rely on a remarkable endometrial somatic stem cells (SSC) population. Here we explore the hypothesis that human endometrial side population (SP) cells correspond to somatic stem cells. We isolated, identified and characterized the SP corresponding to the stromal and epithelial compartments using endometrial SP genes signature, immunophenotyping and characteristic telomerase pattern. We analyzed the clonogenic activity of SP cells under hypoxic conditions and the differentiation capacity in vitro to adipogenic and osteogenic lineages. Finally, we demonstrated the functional capability of endometrial SP to develop human endometrium after subcutaneous injection in NOD-SCID mice. Briefly, SP cells of human endometrium from epithelial and stromal compartments display genotypic, phenotypic and functional features of SSC.

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Section: Methodsmentioning

confidence: 99%

“…The Side Population (SP) method is based on the capability of SP cells to extrude the DNA binding dye Hoechst 33342 via the ATP-binding cassette [24]–[26]. ABC transporters, which include p-glycoprotein and ABCB1, can be specifically blocked by verapamil [27].…”

Section: Introductionmentioning

confidence: 99%

Human Endometrial Side Population Cells Exhibit Genotypic, Phenotypic and Functional Features of Somatic Stem Cells

et al. 2010

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“…Autonomous growth in serum-free media, as non-adherent neurospheres, is thought to be a surrogate marker of neural and malignant progenitors [9], [14], [15], [16], [17], [18]. Identification of normal and tumorigenic stem cells has also been studied by side population analysis in a technique that exploits the increased drug efflux capacity of stem cells [19], [20], [21], [22], [23], [24]. Highly tumorigenic cells often express CD133, a five transmembrane domain glycoprotein also expressed by untransformed hematopoietic and neural progenitors [11], [13], [23], [25].…”

Section: Introductionmentioning

confidence: 99%

Neuroblastoma Cell Lines Contain Pluripotent Tumor Initiating Cells That Are Susceptible to a Targeted Oncolytic Virus

et al. 2009

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BackgroundAlthough disease remission can frequently be achieved for patients with neuroblastoma, relapse is common. The cancer stem cell theory suggests that rare tumorigenic cells, resistant to conventional therapy, are responsible for relapse. If true for neuroblastoma, improved cure rates may only be achieved via identification and therapeutic targeting of the neuroblastoma tumor initiating cell. Based on cues from normal stem cells, evidence for tumor populating progenitor cells has been found in a variety of cancers.Methodology/Principal FindingsFour of eight human neuroblastoma cell lines formed tumorspheres in neural stem cell media, and all contained some cells that expressed neurogenic stem cell markers including CD133, ABCG2, and nestin. Three lines tested could be induced into multi-lineage differentiation. LA-N-5 spheres were further studied and showed a verapamil-sensitive side population, relative resistance to doxorubicin, and CD133+ cells showed increased sphere formation and tumorigenicity. Oncolytic viruses, engineered to be clinically safe by genetic mutation, are emerging as next generation anticancer therapeutics. Because oncolytic viruses circumvent typical drug-resistance mechanisms, they may represent an effective therapy for chemotherapy-resistant tumor initiating cells. A Nestin-targeted oncolytic herpes simplex virus efficiently replicated within and killed neuroblastoma tumor initiating cells preventing their ability to form tumors in athymic nude mice.Conclusions/SignificanceThese results suggest that human neuroblastoma contains tumor initiating cells that may be effectively targeted by an oncolytic virus.

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Isolation and Characterization of Cancer Stem Cells In Vitro

Gedye

Ailles

2012

Methods in Molecular Biology

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The cancer stem cell hypothesis is an appealing concept to account for intratumoral heterogeneity and the observation that systemic metastasis and treatment failure are often associated with the survival of a small number of cancer cells. Whilst in vivo evidence forms the foundation of this concept, in vitro methods and reagents are attractive as they offer opportunities to perform experiments that are not possible in an animal model. While there is abundant evidence that existing cancer cell lines are not reliable models of tumor heterogeneity, recent advances based on well validated novel cancer cell lines established de novo in defined serum-free media are encouraging, particularly in the study of glioblastoma multiforme. In this chapter we wish to broadly outline the process of establishing, characterizing, and managing novel cancer cell lines in defined serum-free media, and discuss the limitations and potential opportunities that may arise from these model systems.

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The Hoechst low-fluorescent profile of the side population: clonogenicity versus dye retention

Cited by 4 publications

References 8 publications

Human Endometrial Side Population Cells Exhibit Genotypic, Phenotypic and Functional Features of Somatic Stem Cells

Human Endometrial Side Population Cells Exhibit Genotypic, Phenotypic and Functional Features of Somatic Stem Cells

Neuroblastoma Cell Lines Contain Pluripotent Tumor Initiating Cells That Are Susceptible to a Targeted Oncolytic Virus

Isolation and Characterization of Cancer Stem Cells In Vitro

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