Sphere Formation Assay for DU145 Cells

Spheres of DU145 cells were formed as previously described [23 (link)]. Briefly, DU145 cells were plated on ultralow attachment culture dishes (Corning, NY, USA) (1 × 10³ cells/well in 6-well plates and 1 × 10⁵ cells/dish in 10 cm dishes) and cultured in DMEM/F-12 (Gibco, NY, USA) supplemented with B27 (Gibco), 4 μg/mL insulin (Sigma, MO, USA), 20 ng/mL epithelial growth factor (EGF; Gibco), and 20 ng/mL basic fibroblast growth factor (bFGF) (ORF, Kopavogur, Iceland) for 10 days at 37 °C and 5% CO₂.

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Saga K., Park J., Nimura K., Kawamura N., Ishibashi A., Nonomura N, & Kaneda Y. (2019). NANOG helps cancer cells escape NK cell attack by downregulating ICAM1 during tumorigenesis. Journal of Experimental & Clinical Cancer Research : CR, 38, 416.

Publication 2019

ultralow attachment culture dishes DMEM/F-12 insulin

Basic fibroblast growth factor Cells Dish Epithelial growth factor Insulin

Corresponding Organization :

Other organizations : Osaka University

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Variable analysis

independent variables

Plating density of DU145 cells (1 × 10^3 cells/well in 6-well plates and 1 × 10^5 cells/dish in 10 cm dishes)

dependent variables

Formation of spheres of DU145 cells

control variables

Culture medium (DMEM/F-12 supplemented with B27, insulin, EGF, and bFGF)
Culture duration (10 days)
Culture conditions (37 °C, 5% CO2)

controls

Positive control: Not specified
Negative control: Not specified

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