Spheroid Culture of Prostate Cancer Cells

Spheroid culture assay was performed as previously described^{24 (link)}. In brief, PCa cells were transfected with NC or miR-628 and plated at a density of 2500 cells/well in triplicates in ultralow attachment six-well culture plates (Corning, St. Louis, MO, USA) and grown in stem cell media consisting of DMEM/F12 (Gibco, Cat# 21331020) media supplemented with B27 (Gibco, Cat#17504044) and N-2 (Gibco, Cat# 17502001). Spheres were visualized and counted 10 days after plating using a light microscope. Sphere area was measured using the Zeiss ZEN lite software.

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Rios-Colon L., Chijioke J., Niture S., Afzal Z., Qi Q., Srivastava A., Ramalinga M., Kedir H., Cagle P., Arthur E., Sharma M., Moore J., Deep G., Suy S., Collins S.P, & Kumar D. (2022). Leptin modulated microRNA-628-5p targets Jagged-1 and inhibits prostate cancer hallmarks. Scientific Reports, 12, 10073.

Publication 2022

ultralow attachment six-well culture plates DMEM/F12 ZEN lite software

Assay Cells Light microscope Stem cell

Corresponding Organization :

Other organizations : North Carolina Central University, Atrium Health Wake Forest Baptist, Georgetown University Medical Center, Georgetown University

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Variable analysis

independent variables

Transfection of PCa cells with NC (negative control) or miR-628

dependent variables

Sphere formation and area

control variables

Cell density (2500 cells/well)
Culture plates (ultralow attachment six-well culture plates)
Culture media (DMEM/F12 media supplemented with B27 and N-2)

controls

Negative control (NC) transfection

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