Colony Formation Assay for Cell Lines

Cells were plated in 6-well culture plates at the concentration of 200 per well. After 24h incubation, cells were treated with PBS, cetuximab, MM-121 or the combination of cetuximab and MM-121 (CM combination) for 10 days to form colonies as previously described (24 (link)). Medium was changed every three days. The colonies were then stained with 0.2% crystal violet with buffered formalin (Sigma). Colony numbers were manually counted using Image J software. Cell numbers ≥50 were considered as a colony.

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Wang D., Qian G., Zhang H., Magliocca K.R., Nannapaneni S., Amin A.R., Rossi M., Patel M., El-Deiry M., Wadsworth J.T., Chen Z., Khuri F.R., Shin D.M., Saba N.F, & Chen Z.G. (2016). HER3 targeting sensitizes HNSCC to cetuximab by reducing HER3 activity and HER2/HER3 dimerization - evidence from cell line and patient derived xenograft models. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(3), 677-686.

Publication 2016

buffered formalin

Cells Cetuximab Crystal violet Formalin Mm 121

Corresponding Organization : Emory University

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Variable analysis

independent variables

Cetuximab
MM-121
Combination of cetuximab and MM-121 (CM combination)

dependent variables

Colony formation

control variables

Cell plating concentration (200 cells per well)
Incubation time (24h)
Colony staining (0.2% crystal violet with buffered formalin)
Colony counting method (Image J software, with colonies ≥50 cells counted)

controls

Positive control not explicitly mentioned
Negative control (PBS treatment)

Annotations

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