Melanoma Tri-Culture Spheroid Generation

Melanoma tri-cultures were generated according to Klicks et al. (2019) (link). Briefly, 1 × 10⁴ cells of CCD-1137SK cells were seeded, followed by simultaneous addition of HaCaT (1 × 10⁴ cells/well) and SK-MEL-28 (ATCC) (2.5 × 10³ cells/well) after three days. For discrimination of individual cell types, HaCaT and SK-MEL-28 cells were labeled with CellTracker Red CMPTX (10 μM) and CellTracker Green CMFDA (10 μM) dye (both Life Technologies), respectively according to manufacturer instructions for 30 min. Tri-culture spheroids were kept in culture for another two days to reach an average diameter of 300 μm.

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Nürnberg E., Vitacolonna M., Klicks J., von Molitor E., Cesetti T., Keller F., Bruch R., Ertongur-Fauth T., Riedel K., Scholz P., Lau T., Schneider R., Meier J., Hafner M, & Rudolf R. (2020). Routine Optical Clearing of 3D-Cell Cultures: Simplicity Forward. Frontiers in Molecular Biosciences, 7, 20.

Publication 2020

SK-MEL-28 CellTracker Red CMPTX CellTracker Green CMFDA

Cells Cmfda Discrimination Melanoma

Corresponding Organization : Mannheim University of Applied Sciences

Other organizations : Central Institute of Mental Health, Heidelberg University, University Hospital Heidelberg, Merck (Germany)

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Variable analysis

independent variables

Cell type and number of cells seeded (1 × 10^4 CCD-1137SK cells, 1 × 10^4 HaCaT cells, and 2.5 × 10^3 SK-MEL-28 cells)

dependent variables

Diameter of tri-culture spheroids (reached an average of 300 μm after two days in culture)

control variables

Labeling of HaCaT and SK-MEL-28 cells with CellTracker dyes (CellTracker Red CMPTX and CellTracker Green CMFDA, respectively)
Culture duration (tri-culture spheroids were kept in culture for another two days)

controls

No positive or negative controls were explicitly mentioned in the provided information.

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