The hepatocellular carcinoma cell line HepG2 used in this study was purchased from American Type Culture Collection (ATCC, Cat. HB-8065™, Rockville, MD, United States). Complete RPMI-1640 medium supplemented with 10% heat-inactivated foetal bovine serum, 1% penicillin-streptomycin (v/v) and 1% L-glutamine (v/v) was used to culture and maintain the cells. All the reagents were purchased from Nacalai Tesque (Kyoto, Japan). Meanwhile, a normal liver cell line (THLE-3) (ATCC) was cultured in Bronchial Epithelial Cell Growth Basal Medium (Lonza, Basel, Switzerland) supplemented with frozen additives without gentamycin/Amphotericin and Epinephrine, 5 ng/mL EGF, 70 ng/mL Phosphoethanolamine and 10% fetal bovine serum. The incubator used for the cell culture work was set at 37 °C with 5% CO2 (Shellab, Cornelius, OR, United States). Upon reaching 80% confluency, the cells were subcultured and transferred into new cell culture flasks. The cells were seeded at a concentration of 1 × 105 cells/mL.
Bronchial epithelial cell growth basal medium
Manufactured by Lonza
Sourced in Switzerland
Bronchial Epithelial Cell Growth Basal Medium is a cell culture medium designed to support the growth and maintenance of primary human bronchial epithelial cells in vitro. It provides the necessary nutrients and growth factors to facilitate the proliferation and differentiation of these specialized cells.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Hepg2, by American Type Culture Collection (1 mentions)
Thle 3, by Lonza (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Hek293t cell, by American Type Culture Collection (1 mentions)
Epidermal growth factor (egf), by Merck Group (1 mentions)
Hct116, by American Type Culture Collection (1 mentions)
Apo transferrin, by Merck Group (1 mentions)
Panc 1, by American Type Culture Collection (1 mentions)
Insulin, by Merck Group (1 mentions)
Hydrocortisone, by Merck Group (1 mentions)
Sodium selenite, by Merck Group (1 mentions)
Coepicm, by ScienCell (1 mentions)
Suit 2, by Health Science Research Resources Bank (1 mentions)
H1975, by American Type Culture Collection (1 mentions)
Beas 2b, by American Type Culture Collection (1 mentions)
H1299, by American Type Culture Collection (1 mentions)
Begm bullet kit, by Lonza (1 mentions)
5 protocols using bronchial epithelial cell growth basal medium
1
Cell Culture: HepG2 and THLE-3 Protocols
2
Generation and Characterization of ACE2/TMPRSS2-Expressing Cell Lines
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HEK293T cells stably expressing human ACE2 and TMPRSS2 were generated by lentiviral transductions as previously described.1 ,16 (link) A highly permissive clone (HAT-24) was identified through clonal selection and used for this study. The HAT-24 line has been extensively cross-validated with the VeroE6 cell line.16 (link) VeroE6-TMPRSS2 (Vero-T) cells were kindly provided by Associate Professor Daniel Watterson (University of Queensland). HAT-24, VeroE6-TMPRSS2 cells were cultured in Dulbecco's Modified Eagle Medium (Gibco, 11995073) containing 10% foetal bovine serum (Gibco, 10099141; DMEM-10%FBS) and VeroE6 cells (ATCC® CRL-1586™) in Minimal Essential Medium (Gibco, 11090099) containing 10% FBS and 1% penicillin-streptomycin (Gibco, 15140122; MEM-10%FBS). pBEC and alveolar epithelial cultures were grown and differentiated until confluent in complete Bronchial Epithelial Cell Growth Basal Medium (Lonza, CC-3171) before use for air liquid interface experiments. All cells were incubated at 37 °C, 5% CO2 and >90% relative humidity. For the Vero-T cell line authentication was performed as previously described.1 ,16 (link) The STR profiling of HAT-24 has been previously described.16 (link) All cell lines tested negative for mycoplasma.
3
Cell Culture Protocols for SARS-CoV-2 Research
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HEK293 cells (ATCC, CRL-1573), HEK293T cells (ATCC, CRL-3216), Vero-E6 cells (ATCC, CRL-1586), and HeLa cells (ATCC-CCL-2) were maintained in Dulbecco’s modified Eagle’s medium (DMEM), A549 cells (ATCC, CCL-185) were maintained in F-12K Nutrient Mixture, supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, and L-glutamine at 37°C in 5% CO2. Calu-3 cells (ATCC, HTB-55) were grown in Eagle’s minimal essential medium supplemented with 10% FBS, 1% penicillin/streptomycin, L-glutamine, and 0.1 mM non-essential amino acids at 37°C in 5% CO2. BEAS-2B cells were obtained from the Conservation Genetics CAS Kunming Cell Bank, and maintained in Bronchial Epithelial Cell Growth Basal Medium (Lonza, Switzerland). Vero-E6 cells and A549 cells expressing human ACE2 (Vero-E6-ACE2 cells, A549-ACE2 cells) were generated by transducing an ACE2-expressing lentiviral vector, and selecting with puromycin, after selecting, cells were subsequently maintained with puromycin.
4
Culturing Human Cancer and Normal Cells
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Human cancer cell lines were grown in McCoy’s 5A Medium (HCT116 [ATCC]), Roswell Park Memorial Institute (RPMI) 1640 (MKN45 [Japanese Cancer Research Resources Bank, Tsukuba, Japan], PANC-1 [ATCC], OE33 [DS Pharma Biomedical Co., Ltd., Osaka, Japan]) or high-glucose Dulbecco’s modified Eagle’s medium (DMEM) (SUIT-2 [Health Science Research Resources Bank, Osaka, Japan]) supplemented with 10% (vol/vol) fetal bovine serum (FBS), 2 mM l -glutamine, 50 U/ml penicillin, and 50 µg/ml streptomycin in a humidified atmosphere containing 5% CO2. Human colorectal epithelial primary (CoEpiC; ScienCell Research Laboratories, Inc., CA, USA), Het1A (non-tumorous esophagus cells, ATCC) and HCEC-1CT (Summit Pharmaceuticals International Corporation, Tokyo, Japan) cells were grown in colonic epithelial cell medium (CoEpiCM; ScienCell), bronchial epithelial cell growth basal medium (Lonza, Basel, Switzerland) and ColoUp medium (DMEM/Medium 199 Earle’s, 4 + 1 (Biochrom Cat# F0435 and Cat# FG0615) containing 4 mM GlutaMAXTM-1 (100×), (Gibco, Cat# 35050-038) 2% cosmic calf serum (Hyclone, Cat# SH30087), 20 ng/ml EGF (Sigma Aldrich, Cat# E9644), 10 μg/ml Insulin (Sigma Aldrich, Cat# I9278), 2 μg/ml Apo-Transferrin (Sigma Aldrich, Cat# T2036), 5 nM sodium-selenite (Sigma Aldrich, Cat# S5261), and 1 μg/ml hydrocortisone (Sigma Aldrich, Cat# H0396)), respectively.
5
In Vitro Culture of Lung Cancer Cell Lines
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Human non-small lung cancer cell lines H1975 (ATCC CRL-5908) and H1299 (ATCC CRL-5803) were obtained from American Type Tissue Collection (ATCC) and maintained in Dulbecco's Modified Eagle Medium (DMEM) (Thermo Fisher Scientific (TFS), cat# 11995-073) containing 10% fetal bovine serum (FBS) (TFS, cat#10437-036). A human bronchial epithelial cell line BEAS-2B (ATCC CRL-9609) was obtained from ATCC and maintained in Bronchial Epithelial Cell Growth Basal Medium (Lonza, CC-3171) containing growth factors and supplements from BEGM BulletKit (Lonza, CC-4175).
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