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Cell culture, generation of palbociclib-resistant cells

LL Lilibeth Lanceta
NL Nadiia Lypova
CO Conor O’Neill
XL Xiaohong Li
ER Eric Rouchka
JC Jason Chesney
YI Yoannis Imbert-Fernandez
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MDA-MB-231 (HTB-26) cells were purchased from the American Type Culture Collection (ATCC) and maintained at 37 °C with 5% CO2. MDA-MB-231 cells were cultured in IMEM (Corning) supplemented with 10% fetal bovine serum (FBS, Invitrogen). MFM-223 cells were purchased from SIGMA and maintained at 37 °C with 5% CO2. MFM-223 cells were cultured in MEM (Corning) supplemented with 10% fetal bovine serum (FBS, Invitrogen). Palbociclib-resistant MDA-MB-231 and MFM-223 cells were established by culturing in media containing palbociclib (0.1–4 μM). Drug was replenished every 3 days. Cells were subcultured every 1–2 weeks with 25% increments in drug concentration. The resistant cells were established after 4–6 months and maintained in the presence of 1 μM palbociclib. Cells were authenticated by the short tandem repeat (STR) assay (Genetica).

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