BEAS-2B (ATCC, Manassas, VA), was cultured in BEGM media (Lonza, Walkersville, MD), with or without 5% fetal bovine serum (FBS) for 8 weeks. All studies were performed using BEAS-2B cells between passage 10 and passage 30, with passage 1 defined as the thawed cells from the supplier. Trypsin-EDTA (0.25%) was used to remove cells from culture flasks for sub-culturing. Cell cultures were incubated in humidified 95% air: 5% CO2. Two million cells were seeded in 75 cm2 culture flasks and sub-cultured at 90% confluence. BEAS-2B cells referred to in this study as “control” were cultured for appropriate passage-matched durations in BEGM without FBS and without arsenite. Cells referred to as “FBS-exposed” were cultured in BEGM, with 5% FBS, without arsenite, for 8 weeks. Cells referred to as “arsenite-exposed” were cultured in BEGM, without FBS, with 1 μM arsenite (Sigma, St. Louis, MO), for 8 weeks. The identity of the BEAS-2B cell line was confirmed using commercial forensic DNA testing based on polymorphic STR markers. BEAS-2B cells were assayed for mycoplasma contamination every two weeks using the MycoAlert system (Lonza, Switzerland). No indication of mycoplasma contamination was found during the course of this study.
Begm media
Manufactured by Lonza
Sourced in United States, Germany, Switzerland
BEGM media is a cell culture medium formulated to support the growth and maintenance of bronchial epithelial cells. It provides the necessary nutrients and growth factors to facilitate the cultivation of these specific cell types.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 medium, by Corning (8 mentions)
H1975, by Corning (6 mentions)
H1975, by Cobioer Biosciences (5 mentions)
Rat tail collagen type 1, by Merck Group (4 mentions)
Antibiotic antimycotic solution, by Wisent (4 mentions)
H1299, by Cobioer Biosciences (4 mentions)
Rpmi 1640 media, by Corning (3 mentions)
H3255, by Lonza (3 mentions)
H1975, by American Type Culture Collection (3 mentions)
Hcc827, by American Type Culture Collection (3 mentions)
Hcc827, by Corning (3 mentions)
Spc a1, by Cobioer Biosciences (3 mentions)
H1650, by Cobioer Biosciences (3 mentions)
Hek293t, by American Type Culture Collection (3 mentions)
Hek293t, by Corning (3 mentions)
Dmem medium, by Corning (3 mentions)
Beas 2b, by American Type Culture Collection (2 mentions)
Type 4 collagen coated 12 well transwell inserts, by Corning (2 mentions)
F 12k nutrient mixture, by Thermo Fisher Scientific (2 mentions)
H2122, by American Type Culture Collection (2 mentions)
28 protocols using begm media
1
Beas-2B Cell Culture and Treatment
2
Bronchial Epithelial Cells from Obese Subjects
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Normal human primary bronchial epithelial (NHBE) cells from non-obese (BMI < 30 kg/m2) and obese (BMI ≥ 30 kg/m2) subjects were purchased from a commercial source (MatTek, MA, United States) 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. Table 1 shows the data from the non-obese and obese subjects. NHBE cells were cultured in BEGM media (Lonza, MD, United States) supplemented with 1% antibiotic antimycotic solution (Wisent, QC, Canada) in tissue culture flasks coated with Type 1 Rat tail collagen (Sigma-Aldrich, Ontario, ON, Canada). Cells were grown to 90% confluency and detached using 0.05% Trypsin-EDTA (Thermo Fisher Scientific, MA, United States). Fifty thousand cells were reserved for RNA extraction.
3
Differentiation of 3D Pseudostratified HBE Cells
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Primary HBE cells were isolated from patients who underwent surgical lung resection for pulmonary diseases in Nanjing Children’s Hospital, as described previously [11 (link)]. HBE cells were plated onto type I and III collagen-coated six-well tissue culture plates and cultured in BEGM media (Lonza, Germany) supplemented with the required additives (Lonza). When the cells reached 80%–90% confluence, traditional monolayer two-dimensional (2D) HBE cells were dissociated using trypsin, and 3 × 105 cells were seeded on type IV collagen-coated 12-well transwell inserts (Costar, ME, USA). The medium was renewed for both the apical and basolateral surfaces every other day. The medium was then changed to air–liquid interface (ALI) medium (BEGM + DMEM + additives) until the HBE cells reached full confluence. After 5 days, the HBE cells were exposed to air, and only the basolateral compartment was cultured in ALI medium. The ALI culture was continued for 4–6 weeks, during which time the cells differentiated into 3D pseudostratified HBE cells. Prior to the experiments, all cultures were maintained at 37 °C in a 5% CO2 incubator.
4
Bronchial Epithelial Cell Culture at ALI
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Primary BECs were revived and expanded in T75 flasks from liquid nitrogen vials using BEGM media (Lonza, Switzerland). Following cell expansion, BECs were trypsinised and seeded in transwell inserts (Corning, United States; 2 × 105 cells per insert) in ALI initial media comprised of bronchial epithelial base medium and Dulbecco's modified eagle medium (BEBM:DMEM; 50:50 ratio) containing hydrocortisone (0.1%), bovine insulin (0.1%), epinephrine (0.1%), transferrin (0.1%), bovine pituitary extract (0.4%) and ethanolamine (80 μM), MgCl2 (0.3 mM), MgSO4 (0.4 mM), bovine serum albumin (0.5 mg/mL), amphotericin B (250 μg/mL), all-trans retinoic acid (30 ng/ml), penicillin/streptomycin (2%), and recombinant human epithelial growth factor (rhEGF) (10 ng/ml) for 3–5 days until confluent. Once confluent, apical media was removed (day 0 of ALI), as previously described (37 (link)). Basal media was changed on alternative days with ALI final media, containing lower rhEGF concentrations (0.5 ng/mL).
5
Culturing Primary Bronchial Epithelial Cells
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Normal human primary bronchial epithelial (NHBE) cells from non-obese and obese subjects were purchased from a commercial source (MatTek, Ashland, MA, 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 1 represents subjects’ characteristics from the non-obese and obese donors. NHBE cells were cultured in BEGM media (Lonza, Walkersville, MD, USA) supplemented with 1% antibiotic antimycotic solution (Wisent, St-Bruno, QC, Canada) in tissue culture flasks coated with Type 1 rat tail collagen (Sigma-Aldrich, Oakville, ON, Canada). Cells were harvested at 90% confluency.
6
Authenticated Cancer Cell Line Cultivation
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The human cancer cell lines H1975, HCC827, H2122, CHO, CHL and H1355 were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA). PC9 cell line was purchased from the Sigma-Aldrich (St. Louis, MO, USA). A549, H3255 were purchased from Cobioer Biosciences CO., LTD (Nanjing, China). H1975, PC-9, HCC827 and EGFR mutant isogenic BaF3 cells were cultured in RPMI 1640 media (Corning, USA) with 10% fetal bovine serum (FBS) and supplemented with 2% L-glutamine and 1% penicillin/streptomycin. A549 and H1355 were cultured in F-12K Nutrient Mixture (kaighn's Modification) (Gibco, USA) with 10% FBS and supplemented with 2% L-glutamine and 1% pen/strep. H3255 was cultured in BEGM media (LONZA, USA) with 10% FBS and supplemented with 2% L-glutamine and 1% pen/strep.
We have authenticated the following cell lines through cell line short tandem repeat (STR) profiling (GENEWIZ, Suzhou, China): H1975, PC9, H3255, HCC827, A549, H2122, H1703. All cell lines matched >90% with lines listed in the ATCC, DSMZ or JCRB Cell Line Bank STR Profile Information.
We have authenticated the following cell lines through cell line short tandem repeat (STR) profiling (GENEWIZ, Suzhou, China): H1975, PC9, H3255, HCC827, A549, H2122, H1703. All cell lines matched >90% with lines listed in the ATCC, DSMZ or JCRB Cell Line Bank STR Profile Information.
7
Culturing Human Cancer Cell Lines
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The human cancer cell lines A549, A431, H3255, H1975, PC-9, HCC827, H23, H460, A549, H358 and H2122 cells were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA). H1975, PC-9, HCC827, H23, H460, H358, H2122 and EGFR mutant isogenic BaF3 cells lines were cultured in RPMI 1640 media (Corning, USA) with 10% fetal bovine serum (FBS) and supplemented with 2% L-glutamine and 1% penicillin/streptomycin. A549 and A431 were cultured in DMEM media (Corning, USA) with 10% FBS and supplemented with 2% L-glutamine and 1% pen/strep. H3255 was cultured in BEGM media (LONZA, USA) with 10% FBS and supplemented with 2% L-glutamine and 1% pen/strep.
8
Cell Line Cultivation and Characterization
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The BEAS-2B cell line was purchased from the Cell Bank of theKunming Institute of Zoology and was cultured in BEGM media (Lonza, CC-3170). Lung cancer cell lines, namely, h1650, A549, SPC-A1, and H1975, were purchased from Cobioer, China with STR documents, and were cultured in RPMI-1640 medium (Corning) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin.
9
Culturing Lung Cancer Cell Lines
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BEAS-2B cell line was purchased from Cell Bank of Kunming Institute of Zoology and cultured in BEGM media (Lonza, Basel, Switzerland; CC-3170). Lung cancer cell lines, including A549, H1299, and SPC-A1, were purchased from Cobioer (Nanjing, China) with short tandem repeat (STR) documents, and were cultured in RPMI-1640 medium (Corning, Manassas, VA, USA) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin.
10
Culturing Virus-Transformed BEAS-2B Cells
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The human virus-transformed bronchial epithelial cell line BEAS-2B was purchased from ATCC and cultured in BEGM media (Lonza), supplemented with a SingleQuot kit (Lonza) and antibiotic-antimycotic solution (Invitrogen) in a 5% CO2 incubator. All culture containers were pre-coated with 30 μg/ml PureCol (Advanced BioMatrix).
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