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Longer uninterrupted dosing is not recommended because receptor sensitivity can diminish without cycling

PMID: 17369935 Sudakov KV, Umriukhin PE, Rayevsky KS

The hAMY 3 R cells were cultured in growth medium consisting of DMEM (Gibco, #31966-021) supplemented with 10% v/v FBS (Gibco, #16140-071), 1% v/v Penicillin-Streptomycin (Gibco, #15140-122), 0.5 mg/mL Geneticin G418 (Gibco, #10131-027), 0.4 mg/mL Hygromycin (Invitrogen, #10687010) and 250 nM Methotrexate (Sigma, A6770)