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Non adherent plates

Manufactured by Corning

Non-adherent plates are a type of cell culture plate designed to prevent the attachment and growth of cells on the surface. These plates are typically made of a specialized material or treated with a coating that inhibits cell adhesion. They are commonly used in applications where the preservation of cell suspension or prevention of cell attachment is desired, such as in the culture of suspension cells, stem cells, or certain types of primary cells.

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3 protocols using non adherent plates

1

2D and Mammosphere Growth Assay

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Cells were seeded in triplicate in 12 well plates, harvested at 24, 48, 72 and 96 hours and counted using the CASY TT cell counter (Roche) for 2D growth. For mammosphere cultures, cells were dissociated enzymatically (Trypsin) and mechanically by pipetting to single-cell suspension and plated on non-adherent plates (Corning) at a concentration of 1000 cells/ml. Cells were grown in a serum-free RPMI 1640 with Pen/strep and L-Glut, supplemented with B27 (Invitrogen), 20 ng/ml EGF (Sigma), 20 ng/ml b-FGF (Sigma), BSA 0.4% and 4 μg/ml Heparin (Sigma). Mammospheres were grown for 7 days and colonies counted under a bright field microscope.
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2

Isolation and Maintenance of HGSOC Spheroids

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Ascites fluid from HGSOC patients was passed through a 40 μm nylon mesh and washed with PBS to remove single cells. Isolated spheroids were maintained in non-adherent plates (Corning).
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3

Quantifying Cancer Stem Cell Spheres

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Five hundred cancer cells were seeded on non-adherent plates (Corning) in serum-free media. After 7 days, colonies greater than 30 cells were counted. Primary spheres were dissociated with trypsin and single cells passaged for second and third passage. In some experiments, single cells were embedded in Matrigel (BD Biosciences) as described.54 (link) Cells were washed with 1 × phosphate-buffered saline and fixed with 100% MetOH for 10 min at room temperature. Crystal violet stain (Fluka AG 61135, 0.1% in 5% EtOH, Sigma-Aldrich, St Loise, MO, USA) was used to stain colonies prior to counting. Quantification of mammosphere and tumorsphere numbers was accomplished using a Multisizer 3 Coulter Counter (Beckman-Coulter, Brea, CA, USA) that provide number and size distribution of particles between 40 and 336 μm.
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