The formation of tumorspheres was used to evaluate the self-renewal ability of melanoma-initiating cells. In brief, melanoma cells were suspended in a stem cell medium and plated at a density of 5,000 cells/well in Corning® ultralow adherent six-well dishes (Corning Incorporated, Corning, NY, USA). The stem cell medium contained DMEM/F12, B27 supplement (1×), ITS (1×), EGF, and bFGF (both at a concentration of 20 ng/mL). The cells were cultured in the stem cell medium for 7 days, and then tumorspheres were counted under a conventional microscope. For second passage tumorspheres, those of the first passage were washed with PBS, dissociated with a cell dissociation reagent (StemPro® Accutase®; Thermo Fisher Scientific, Waltham, MA, USA), and then propagated.
Ultralow adherent six well dishes
Manufactured by Corning
Sourced in United States
The Ultralow adherent six-well dishes are a specialized lab equipment product designed for cell culture applications. These dishes provide a low-attachment surface that minimizes cell adhesion, promoting the formation of 3D cell aggregates or spheroids. The dishes feature six individual wells, allowing for multiple experimental conditions or cell types to be cultured simultaneously.
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Lab products found in correlation
2 protocols using ultralow adherent six well dishes
1
Evaluating Melanoma-Initiating Cells' Self-Renewal
2
Lung Cancer Stem Cell Isolation
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Tumor sphere-forming assays are used to identify CSCs, based on their capacity to realize differentiation and self-renewal at the level of single cells [26 (link)]. Briefly, lung cancer cells (3000 cells/well) were cultured in ultra-low adherent six-well dishes (Corning, Tewksbury, MA). The cells were suspended in stem cell medium composed of DMEM/F12 (Dulbecco’s modified Eagle medium/Ham’s F-12 medium), with 1 × B27, 1 × ITS, 20 ng/mL EGF, and 20 ng/mL bFGF. The number of tumor spheres was counted after 7 days. For the formation of the second-passage tumor spheres, tumor spheres were dissociated by the StemPro® Accutase® Cell Dissociation Reagent and propagated.
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