Commercial lung cell lines BEAS-2B [28 (link)] (CRL-9609, a human bronchial epithelial cell line) and HFL1 [29 (link)] (CCl-153, a human fetal lung fibroblast cell line) were purchased from ATCC (Virginia, USA). The cell lines were grown as per the supplier’s recommendations. BEAS-2B and HFL1 were cultured in RPMI and DMEM, respectively, supplemented with antibiotics and 10% FCS and incubated in 5% CO2.
Crl 9609
Manufactured by American Type Culture Collection
Sourced in United States
CRL-9609 is a cell line product offered by the American Type Culture Collection. It is a cell line derived from human skin fibroblasts. The core function of this product is to provide a standardized cell culture for research and experimental purposes.
Automatically generated - may contain errors
Lab products found in correlation
Beas 2b, by American Type Culture Collection (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Crl 1730, by American Type Culture Collection (2 mentions)
Penicillin g, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Htb 77, by American Type Culture Collection (2 mentions)
Trypan blue, by Merck Group (2 mentions)
Trypsin edta solution, by Merck Group (2 mentions)
Ccl 153, by American Type Culture Collection (1 mentions)
Lhc 9 medium, by Thermo Fisher Scientific (1 mentions)
L2020, by Merck Group (1 mentions)
Poly hema, by Merck Group (1 mentions)
Ccl 34, by American Type Culture Collection (1 mentions)
Rpmi 1640 glutamax 1, by Thermo Fisher Scientific (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Crl 2110, by American Type Culture Collection (1 mentions)
Mle 12, by American Type Culture Collection (1 mentions)
Ccl 185emt, by American Type Culture Collection (1 mentions)
L dmem, by Thermo Fisher Scientific (1 mentions)
21 protocols using crl 9609
1
Culturing lung cell lines BEAS-2B and HFL1
2
Culturing Human Bronchial Epithelial Cells
3
Culturing and Coating Protocols for Glioma and Control Cell Lines
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Human glioma cell lines (LN229, LN319, LN444, LN751, SF188) [25 (link),26 (link),27 (link)], human normal lung bronchial epithelial BEAS-2B cells (CRL-9609, ATCC, Manassas, VA, USA), and human embryonic kidney 293T cells were cultured on adhesive cell culture dishes in DMEM medium (10-013-CV, CORNING, Corning, NY, USA) supplemented with 10% fetal bovine serum (FBS; 10437-028, Thermo Fisher Scientific, Waltham, MA, USA) and 1% Penicillin and Streptomycin (SV30010, Cytiva, Marlborough, MA, USA) under an atmosphere of 95% air and 5% CO2 at 37 °C. Colorectal cancer cell line HCT116 was maintained in McCoy’s 5A Medium with 10% FBS. All cell lines were authenticated via short tandem repeat profiling and were regularly tested for mycoplasma.
For laminin experiments, cell culture-treated plates (TPN1006, Alkali Scientific, Fort Lauderdale, FL, USA) were coated with 10 or 50 µg/mL of laminin (L2020, Sigma-Aldrich, Burlington, MA, USA) according to the manufacturer’s protocol.
For non-adhesive cell culture conditions, non-cell culture-treated plates (351143, Corning) were coated with hydrogel (Poly-HEMA; poly 2-hydroxyethyl methacrylate, P3932, Sigma-Aldrich). Poly-HEMA (20 mg/mL dissolved in 95% ethanol on a rotating wheel at 42 °C for 24 h) was added to the plates and dried overnight. The number of viable cells was determined by performing the trypan blue dye exclusion test.
For laminin experiments, cell culture-treated plates (TPN1006, Alkali Scientific, Fort Lauderdale, FL, USA) were coated with 10 or 50 µg/mL of laminin (L2020, Sigma-Aldrich, Burlington, MA, USA) according to the manufacturer’s protocol.
For non-adhesive cell culture conditions, non-cell culture-treated plates (351143, Corning) were coated with hydrogel (Poly-HEMA; poly 2-hydroxyethyl methacrylate, P3932, Sigma-Aldrich). Poly-HEMA (20 mg/mL dissolved in 95% ethanol on a rotating wheel at 42 °C for 24 h) was added to the plates and dried overnight. The number of viable cells was determined by performing the trypan blue dye exclusion test.
4
BEAS-2B Cell Culture Protocol
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The human bronchial epithelial cell line, BEAS-2B, transformed with an adenovirus 12-SV40 virus hybrid (CRL-9609; ATCC, Manassas, VA, USA), was cultured in DMEM/F12 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS), penicillin (100 U/mL), and streptomycin (100 μg/mL).
5
BEAS-2B Cell Culture and Optimization
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BEAS-2B (human bronchial epithelial cell line) (ATCC® CRL9609™) was cultured with Bronchial Epithelial Cell Growth Basal Medium (BEBM) in tissue culture flasks coated with 0.01 mg/mL fibronectin, 0.03 mg/mL bovine collagen type I, and 0.01 mg/mL bovine serum albumin (BSA). The cells were incubated at 37°C in a humidified environment with 5% CO2 [50 (link)].
6
Measurement of IFITM1 and Interferon in Cell Lines
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Human umbilical vein endothelial cells (HUVECs, CRL-1730, ATCC), human bronchus epithelial cells (BEAS-2Bs, CRL-9609, ATCC) were used for detecting the expression of IFITM1 and secretion of interferon-ɑ/β. Madin Darby canine kidney cells (MDCK, CCL-34, ATCC) were used for plaque assay. These cell lines were cultured in DMEM (Gibco) supplemented with 10% FBS (Gibco), 100 U/mL of penicillin G and 100 μg/mL of streptomycin (Gibco) at 37 °C in a 5% CO2 incubator. The cells in passages 5~8 were seeded into six-well plates and cultured for 24 h before each experiment.
7
Pathogenic Bacteria Cultivation and Cell Line Maintenance
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S. aureus strains MW2, a highly pathogenic and a multidrug-resistant strain was obtained from Network on Antimicrobial Resistance in Staphylococcus aureus (NARSA). Streptococcus pyogenes M1T15448 was obtained as described previously37 . Staphylococcus strain was grown in tryptic soy broth (TSB) or TS agar. Streptococcus strain was grown in Todd-Hewitt broth supplemented with 0.5% yeast extract or 5% sheep-blood TS agar. Escherichia coli strain (DH5α, BL21 DE3-pLysS) were grown in Luria-Bertani (LB) or LB agar with or without ampicillin (100 µg/ml). All bacterial culture media were obtained from Difco (BD and Co, Sparks, MD). Antibiotics unless otherwise indicated were used at the following concentrations: 100 μg/ml of ampicillin and 10 μg/ml of chloramphenicol for E. coli, 10 μg/ml of erythromycin and 10 μg/ml of chloramphenicol and 1.5 μg/ml of anhydrotetracycline for S. aureus. Human pharyngeal carcinoma cell line (Detroit 562, CCL-138ATCC), and BEAS-2B (normal lung/bronchoalveolar immortalized cell lines, CRL-9609, ATCC), were grown in 24 well plates to obtain a confluent culture. Detroit 562 cell lines were grown in RPMI 1640-GlutaMax-I (GIBCO) tissue culture medium with 10% fetal bovine serum. BEAS-2B cell lines were grown in a serum-free medium, LHC-9, supplemented with epidermal growth factor and pituitary extract.
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S. aureus strains MW2, a highly pathogenic and a multidrug-resistant strain was obtained from Network on Antimicrobial Resistance in Staphylococcus aureus (NARSA). Streptococcus pyogenes M1T15448 was obtained as described previously37 . Staphylococcus strain was grown in tryptic soy broth (TSB) or TS agar. Streptococcus strain was grown in Todd-Hewitt broth supplemented with 0.5% yeast extract or 5% sheep-blood TS agar. Escherichia coli strain (DH5α, BL21 DE3-pLysS) were grown in Luria-Bertani (LB) or LB agar with or without ampicillin (100 µg/ml). All bacterial culture media were obtained from Difco (BD and Co, Sparks, MD). Antibiotics unless otherwise indicated were used at the following concentrations: 100 μg/ml of ampicillin and 10 μg/ml of chloramphenicol for E. coli, 10 μg/ml of erythromycin and 10 μg/ml of chloramphenicol and 1.5 μg/ml of anhydrotetracycline for S. aureus. Human pharyngeal carcinoma cell line (Detroit 562, CCL-138ATCC), and BEAS-2B (normal lung/bronchoalveolar immortalized cell lines, CRL-9609, ATCC), were grown in 24 well plates to obtain a confluent culture. Detroit 562 cell lines were grown in RPMI 1640-GlutaMax-I (GIBCO) tissue culture medium with 10% fetal bovine serum. BEAS-2B cell lines were grown in a serum-free medium, LHC-9, supplemented with epidermal growth factor and pituitary extract.
8
Culturing Normal Lung Epithelial Cells
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Human-derived lung normal epithelial cells (BEAS-2B, CRL-9609, ATCC) and Mouse-derived lung normal epithelial cells (MLE-12, CRL-2110, ATCC) were maintained in 10% foetal bovine serum (Gibco, Grand Island, NY, USA), supplemented with 1% penicillin and 100 mg/mL streptomycin and grown at 5% CO2 and 37 °C.
9
ROCK1 Inhibition in NSCLC Cell Lines
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The normal pulmonary epithelial cell line BEAS-2B (American Type Culture Collection (ATCC)® no. CRL-9609™) and the NSCLC cell line A549 (ATCC® no. CCL-185EMT™) were purchased from ATCC (Manassas, VA, USA). The cells were cultured in Low Glucose-Dulbecco's modified Eagle's medium (L-DMEM; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) containing low glucose, 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.) and 1% streptomycin and penicillin at 37°C in an atmosphere containing 5% CO2. To inhibit ROCK1 activity, Y-27632 (5 mM; cat. no. S1049; Selleck Chemicals, Houston, TX, USA) was added to the medium for 4 h (21 (link)).
10
Viral Interaction with Mammalian Cell Lines
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Human umbilical vein endothelial cells (HUVECs, CRL-1730, ATCC), human bronchus epithelial cells (BEAS-2Bs, CRL-9609, ATCC) and Madin-Darby canine kidney (MDCK, CCL-34, ATCC) are commonly used cell lines for evaluation of interactions between viruses and mammalian cells. In this study, HUVECs and BEAS-2Bs were used for detecting IFIT1 expression and IFN-α/β secretion. MDCK cells were used for the plaque assay. HUVECs, BEAS-2Bs and MDCK were cultured in DMEM medium (Gibco, USA) supplemented with FBS 10% (Gibco, USA), penicillin G 100 U/ml (Gibco, USA) and streptomycin 100 g/mL (Gibco, USA) in a CO2 5% incubator at 37°C. The cells 5–8 passages were seeded into six-well plates and cultured for 24 h before each experiment.
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