To generate ACI23 and OV90 spheroids, 500 cells/well in 50 μL were seeded in TIC-enriching conditions (stem cell media in ultra-low attachment flat bottom 96 well plates (Corning)). After 7 days in culture 3 fields per well were imaged at 10× magnification using Axio Observer A1 Inverted Microscope and analyzed using ImageJ Software 1.48v. Spheroids with diameter ≥50 microns were quantified. Spheroid efficiency was calculated as spheroids ≥50 microns/cells per well).
Ultra low attachment flat bottom 96 well plates
Manufactured by Corning
Sourced in United States
Ultra-low attachment flat bottom 96 well plates are a type of lab equipment designed to inhibit cell attachment. They feature a flat bottom and a specialized surface treatment that minimizes cell adhesion, promoting the growth of suspension cultures and the formation of spheroids.
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Lab products found in correlation
3 protocols using ultra low attachment flat bottom 96 well plates
1
Spheroid Formation from Ovarian Cancer Cells
2
Sphere Forming Assay for Stemness
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The sphere forming assay was performed by seeding single cell suspensions containing 100 cells per well in ultra-low attachment flat bottom 96-well plates (Corning, NY, USA) for 2 wk. All cells were maintained in a defined serum-free medium, DMEM/F12 (Lonza), supplemented with recombinant human epidermal growth factor (EGF, 20 ng/mL, Gibco), basic fibroblast growth factor (bFGF, 20 ng/mL, Gibco), and N-2 Supplement (Gibco), which was changed every 3 d. Subsequently, the spheres larger than 50 μm were identified, and 24 wells were evaluated for each cell culture.
3
Sphere formation assay in A2780-SP cells
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A2780-SP cells were plated in Corning ultra-low attachment flat-bottom 96-well plates at a density of 6000 viable cells per well and grown in CSC medium. After 24 h, A2780-SP cells were treated with the compound. Sphere cells were grown for 60 h. After 60 h, we analyzed the sphere-forming confluence using IncuCyte (BioTek, Winooski, VT, USA).
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