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2 protocols using bronchial epithelium basal medium bebm

1

Cell Line Culturing and Transfection

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The SK-Hep1, Hep3B, Huh-7, HepG2 and SNU449 cell lines were obtained from the Korean Cell Line Bank (Korea). SH-J1 cells were provided by Dr. Dae-Ghon Kim (Medical School, Chonbuk National University) (Kim et al., 2002 (link)). HEK293FT and THLE-2 cells were purchased from Invitrogen and American Type Culture Collection (USA), respectively. HEK293FT and liver cancer cells were cultured at 37°C under 5% CO2 in DMEM supplemented with 10% FBS, 100 units/ml of penicillin and 100 μg/ml streptomycin. THLE-2 cells originated from human primary normal liver cells were plated on culture plates pre-coated with a solution containing 0.01 mg/ml fibronectin, 0.03 mg/ml bovine collagen type I and 0.01 mg/ml bovine serum albumin dissolved in Bronchial Epithelium Basal Medium (BEBM, Lonza). THLE-2 cells were cultured at 37°C under 5% CO2 in BEBM supplemented with Bronchial Epithelium Cell Growth Medium (BEGM) SingleQuots (Lonza). Lipofectamine 2000 (Invitrogen) was used to transfect expression vectors into cells. The same control, VRK1, VRK3 and p53 expression vectors were used as described previously (Kang and Kim, 2006 (link); Kang et al., 2008 (link); Lee et al., 2015b (link)).
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2

Bronchial Epithelial Cell Culture Protocol

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Insulin, bovine serum albumin, fetal bovine serum (FBS), bovine hypothalamus acetone power, hydrocortisone, recombinant human epidermal growth factor, epinephrine hydrochloride, fibronectin, triiodothyronine, transferrin, trans retinoic acid, trypsin, gentamicin, were obtained from Sigma (St. Louis, MO, USA). Bronchial epithelium basal medium (BEBM®) was purchased from LONZA (Clonetics®, Walkersville, MD, USA).
Penicillin G, RPMI-1640 media, streptomycin sulfate, amphotericin B and L-glutamine were purchased from Invitrogen (Melbourne, VIC, Australia). Ultroser G was supplied from Ciphergen (Cergy-Saint-Christophe, France). Collagens type I and all tissue culture plastic ware were purchased from BD (Bedford, MA, USA).
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