Trypsin/EDTA and antibiotic/antimycotic solution (Penicillin/Streptomycin, thereafter P/S) were from Life Technologies (Carlsbad, CA). Fetal bovine serum (FBS) was from Omega Scientific (Tarzana, CA). DMEM-F12 was from ATCC (Mannasas, VA) and DMEM from Corning (Manassas, VA). BEBM basal medium and BEGM supplements and growth factors were purchased from Lonza (Walkersville, MD). Anti-human fibronectin (mouse ant-human ascites fluid, clone IST-4), α-smooth muscle actin (mouse anti-human clone 1A4), β-tubulin (mouse anti-human ascites, clone TUB 2.1), and budesonide were from Sigma (St. Louis, MO). Recombinant human LIGHT/TNFSF14, lymphotoxin α1/β2, TNF and TGF-β1, human antibodies to MMP-9 and Vimentin, and Human active MMP-9 Flurokine E kit, TGF-β1 DuoSet ELISA, and CCL5/RANTES Quantikine ELISA were obtained from R&D Systems (Minneapolis, MN). Recombinant human LIGHT FLAG-tagged was from Enzo Life Sciences (Farmingdale, NY). Anti-human CD270 (HVEM clone eBioHVEM-122), CD54 (ICAM-1, clone HA58), and CD324 (E-Cadherin, clone DECMA-1) were from eBioscience (San Diego, CA). Anti-human CD106 (VCAM-1, clone STA) and LTβR (clone 31G4D8) were from BioLegend (San Diego, CA). Human anti-E-Cadherin (rabbit anti-human, H-108) was from Santa Cruz Biotecnhology, Inc (Dallas, TX).
Bebm basal medium
Manufactured by Lonza
Sourced in United States
BEBM basal medium is a cell culture medium designed to support the growth and maintenance of various cell types. It provides a balanced formulation of essential nutrients, vitamins, and salts to support cell viability and proliferation. The core function of BEBM basal medium is to create a suitable environment for cell culturing and experimentation.
Automatically generated - may contain errors
Lab products found in correlation
Budesonide, by Merck Group (1 mentions)
Dmem f12, by American Type Culture Collection (1 mentions)
Tgf β1 duoset elisa, by R&D Systems (1 mentions)
Ccl5 rantes quantikine elisa, by R&D Systems (1 mentions)
Recombinant human tnfsf14 light, by R&D Systems (1 mentions)
Lymphotoxin α1 β2, by R&D Systems (1 mentions)
Ltβr clone 31g4d8, by BioLegend (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Omega Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Corning (1 mentions)
Dmem medium, by Merck Group (1 mentions)
Opti mem medium, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Begm singlequots kit, by Lonza (1 mentions)
Sars cov 2 spike protein s1 s2, by Sino Biological (1 mentions)
Begm media, by Lonza (1 mentions)
Rat tail collagen type 1, by Merck Group (1 mentions)
Antibiotic antimycotic solution, by Wisent (1 mentions)
Human tgf β1, by R&D Systems (1 mentions)
Thle 3 cells, by American Type Culture Collection (1 mentions)
6 protocols using bebm basal medium
1
Cytokine and Extracellular Matrix Signaling
2
CCA Cell Lines and Culture Conditions
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Human CCA cell lines SG231, CCLP1, HuCCT1, TFK1 and 1 noncancerous cholangiocyte cell line (H69) were got from Japanese Cancer Research Resources Bank (Ibaraki City, Japan). SG231 cells were kept in OPTI-MEM medium (Invitrogen, Carlsbad, CA, USA) with 5% fetal bovine serum (FBS, HyClone, Logan, UT, USA). CCLP1 cells were kept in DMEM medium (Sigma, St. Louis, MO, USA) with 10% FBS (HyClone). Besides, TFK1 and HuCCT1 cells were kept in RPMI 1640 medium (Invitrogen) with 10% FBS (HyClone). H69 cells were kept in BEBM Basal Medium (Lonza, Basel, Switzerland) adding with growth elements (BEGM SingleQuot Kit) with 10% FBS (HyClone). All cells were kept in a wet 37°C environment with 5% CO2. Experiments were conducted when cells reached 80% confluence and were performed in “serum-free medium” (SFM).
3
BEAS-2B Cell Culture Conditions
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Human BEAS-2B cells (American Type Tissue Collection) were plated in six-well plates at a density of 1.5-2x106 cells/well, cultured in complete BEAS-2B culture medium (BEBM) and incubated at 37˚C, 5% CO2. Complete BEBM was formulated by supplementing BEBM basal medium (Lonza Group Ltd.) with a BEGM SingleQuots kit (Lonza Group Ltd.), which consisted of bovine pituitary extract, hydrocortisone, human epidermal growth factor, epinephrine, transferrin, insulin, retinoic acid, triiodothyronine, gentamicin and amphotericin-B.
4
NHBE Cell Response to SARS-CoV-2 Spike Protein
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Normal human primary bronchial epithelial (NHBE) cells from non-obese and obese subjects were purchased from a commercial source (MatTek, MA, USA and ATCC, VA, USA) or obtained from the Biobank of the Quebec Respiratory Health Research Network at the Meakins-Christie Laboratories, Research Institute of the McGill University Health Centre (GLEN site). Table S6 shows the data from the non-obese and obese subjects. NHBE cells were cultured in BEGM media (Lonza, MD, USA) supplemented with 1% antibiotic antimycotic solution (Wisent, QC, CA) in tissue culture 12 well plates coated with Type 1 Rat tail collagen (Sigma-Aldrich, Ontario, Canada). Cells were grown to 90% confluency and starved using BEBM Basal Medium (Lonza, MD, USA) supplemented with 1% antibiotic antimycotic solution (Wisent, QC, CA) over night. The next day cells were stimulated with 1ug/ml of SARS-CoV-2 spike protein (S1+S2) (Sino Biological Inc., Beijing, China) for 3h. Culture media was collected in microtubes, centrifuged 5000g for 5 min. Supernatants were aliquoted into fresh microtubes and frozen at -80°C.
5
Analyzing C-82 effects on hepatocytes and stellate cells
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C-82, the active form of PRI-724, was kindly provided by PRISM BioLab (Yokohama, Japan). The effect of C-82 in hepatocytes was analyzed using THLE-3 cells (CRL-3583, ATCC, USA), cultured in BEBM basal medium (CC-3171, Lonza, Switzerland) with 10% FBS, whereas the effect of C-82 in hepatic stellate cells was analyzed using the LX-2 Human Hepatic Stellate Cell Line (Sigma-Aldrich, Saint Louis, USA), cultured in DMEM with 2% FBS. LX-2 cells were exposed to human TGF-β1 (0.5 ng/ml) (R&D Systems, Minneapolis, USA) for 24 h before use. After a 6-h exposure to 0.5 μM C-82, total RNA was extracted using the miRNeasy Mini Kit (Qiagen).
6
Culturing NSCLC and Bronchial Cell Lines
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Human NSCLC cell lines (A549, H1299, H23, H460, and H1792) were maintained in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum (FBS; Life Technologies, Grand Island, NY, USA). BEAS-2B human bronchial epithelial cells were grown in BEBM basal medium (Lonza, Walkersville, MD, USA) with 10% FBS in a humidified incubator at 37 °C with 5% CO2. These cell lines were validated using the short tandem repeat profiling technique.
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