Culture methods and target cells are summarized in Table 1 . BEAS-2B (No. CRL-9609; ATCC, Manassas, VA) immortalized human lung epithelial cells were cultured in BEGM media with a Single Quot supplement kit (No. CC-3170; Lonza, Basel, Switzerland) at 37°C in 5% CO2. LNCaP (No. CRL-1740; ATCC) human prostate adenocarcinoma cells were cultured using phenol-red free RPMI-1640 (No. 11835; Life Technologies, Grand Island, NY) with 10% fetal bovine serum (FBS) and penicillin/streptomycin at 37°C in 5% CO2. PLHC-1 (No. CRL-2406; ATCC) fish (Poeciliopsis lucida) hepatoma cells were cultured using EMEM media (No. 30–2003; ATCC) with 5% FBS and penicillin/streptomycin at 30°C in 5% CO2.
Single quot supplement kit
Manufactured by Lonza
Sourced in Switzerland
The Single Quot supplement kit is a laboratory equipment designed to provide a standardized supplementation for cell culture media. It contains a pre-measured set of essential nutrients and factors required for optimal cell growth and proliferation. The kit offers a convenient and consistent way to supplement cell culture environments.
Automatically generated - may contain errors
Lab products found in correlation
Phenol red free rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Lncap, by American Type Culture Collection (1 mentions)
Beas 2b, by American Type Culture Collection (1 mentions)
Plhc 1, by American Type Culture Collection (1 mentions)
Emem media, by American Type Culture Collection (1 mentions)
Recombinant human egf, by Thermo Fisher Scientific (1 mentions)
Pd98059, by MedChemExpress (1 mentions)
Gefitinib, by MedChemExpress (1 mentions)
2 protocols using single quot supplement kit
1
Culture Methods for Cell Lines
2
Lung Cancer Cell Culture and EGFR Signaling
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A549, H1299, and H460 human lung cancer cells were maintained in RIPM-1640 medium containing 10% fetal bovine serum (FBS), 100 U/mL penicillin and 0.1 mg/mL streptomycin. BEAS-2B cells were maintained in bronchial epithelial growth medium (BEGM) (Solarbio, Beijing, China) with growth factors, cytokines and supplements (BR, Lonza, Switzerland) in the SingleQuot® supplement kit. Cells were maintained at 37 °C in a humidified atmosphere with 5% CO2. All cell lines were recently authenticated by cellular morphology and short tandem repeat analysis at Microread Inc. (Beijing, China).
For the activation of EGFR/ERK signaling, H1299 cells were starved overnight and incubated with human recombinant EGF (Invitrogen, Carlsbad, CA, USA) for 0–48 h or treated with 0–100 ng/mL EGF for 5 min or 48 h. For inhibition of EGFR/ERK signaling, cells were pre-incubated with 10 μM gefitinib (MedChem Express, New Jersey, USA) or 20 μM PD98059 (MedChem Express, New Jersey, USA) for 1 h and then exposed to 50 ng/ml EGF for 5 min or 48 h.
For the activation of EGFR/ERK signaling, H1299 cells were starved overnight and incubated with human recombinant EGF (Invitrogen, Carlsbad, CA, USA) for 0–48 h or treated with 0–100 ng/mL EGF for 5 min or 48 h. For inhibition of EGFR/ERK signaling, cells were pre-incubated with 10 μM gefitinib (MedChem Express, New Jersey, USA) or 20 μM PD98059 (MedChem Express, New Jersey, USA) for 1 h and then exposed to 50 ng/ml EGF for 5 min or 48 h.
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