Cell lines A549, NCI-H1975 and NCI-H1650 were purchased from the American Type Culture Collection (ATCC, Molsheim Cedex, France). In addition, the Lewis Lung carcinoma (3LL) cell line (gift from Dr. Carsten Riether, Department of Clinical Research, University of Bern; derived from the lung of a C57BL/6J mouse) was included. A549 and 3LL were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Life Technologies, 10938-025, Merelbeke, Belgium) supplemented with 10% FBS (Life Technologies, 10270-106), 1% penicillin (100 U/mL) /streptomycin (100 µg/mL; Life Technologies, 15140-122) and 2 mM l -glutamine (l -Glut, Life Technologies, 25030-024). NCI-H1975 and NCI-H1650 were cultured in Roswell Park Memorial Institute Medium (RPMI, Life Technologies, 52400-025) supplemented as described above. Cells were grown as monolayers and were maintained in exponential growth in 5% CO2 + 95% air in a humidified incubator at 37 °C. All cell cultures were confirmed as Mycoplasma free using the Mycoalert® Mycoplasma detection kit (Lonza, LT07-218, Verviers, Belgium).
Nci h1650
Manufactured by Thermo Fisher Scientific
Sourced in United States
The NCI-H1650 is a human lung adenocarcinoma cell line. It is a commonly used in vitro model for cancer research.
Automatically generated - may contain errors
Lab products found in correlation
Nci h1650, by American Type Culture Collection (8 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Nci h1975, by Thermo Fisher Scientific (5 mentions)
Nci h1975, by American Type Culture Collection (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Nci h1299, by Thermo Fisher Scientific (4 mentions)
Mycoalert mycoplasma detection kit, by Lonza (3 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions)
Nci h460, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640, by Thermo Fisher Scientific (3 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (2 mentions)
Hcc827, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Nci h358, by Thermo Fisher Scientific (2 mentions)
Beas 2b, by American Type Culture Collection (2 mentions)
Nci h1299, by American Type Culture Collection (2 mentions)
L glut, by Thermo Fisher Scientific (1 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
13 protocols using nci h1650
1
Cell Culture of Cancer Cell Lines
2
NSCLC Cell Culture Protocol
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The human NSCLC cell lines NCI-H1975, NCI-H1650, HCC827, LUDLU-1 and A549 were purchased from the American type cell culture collection (ATCC, Rockville MD, USA). The A549 cell line was cultured in DMEM supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin and 1% L-glutamine (Life Technologies, Merelbeke, Belgium). NCI-H1975, NCI-H1650, HCC827 and LUDLU-1 cells were cultured in RPMI supplemented as described above with addition of 1mM sodium pyruvate (Life Technologies). Cells were grown as monolayers and were maintained in exponential growth at 5% CO2/95% air in a humidified incubator at 37°C to obtain normoxic conditions and in a humidified Bactron IV anaerobic chamber (Shel Lab, OR, USA, 1% O2, 5% CO2, 95% N2) to obtain hypoxic conditions. Cells used for experiments in hypoxic conditions were first grown overnight under normoxia to allow the cells to attach to the bottom. Cell cultures were regularly tested for absence of mycoplasma using the Mycoalert® Mycoplasma detection kit (Lonza, Verviers, Belgium) .
3
Culturing and Transducing Cell Lines
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Cell lines A549, NCI-H1975 and NCI-H1650 were purchased from the American Type Culture Collection (ATCC). In addition, the Lewis Lung carcinoma (LLC) cell line (gift from Dr. Carsten Riether, Department of Clinical Research, University of Bern; derived from the lung of a C57BL/6J mouse) was transduced with the viral vector pCH-EF1a-MmCD70-Ires-eGFP-T2A-Puro (Leuven Viral Vector Core, KU Leuven, Belgium) to stably express CD70 (Suppl. Figure S3) A549 and LLC were cultured in Dulbecco’s Modified Eagle Medium (DMEM, 10938–025, Life Technologies) supplemented with 10% FBS (10270–106, Life Technologies), 1% penicillin (100 U/mL)/streptomycin (100 µg/mL; 15140–122, Life Technologies) and 2 mM L-glutamine (L-Glut, 25030–024, Life Technologies). NCI-H1975 and NCI-H1650 were cultured in Roswell Park Memorial Institute Medium (RPMI, 52400–025, Life Technologies) supplemented as described above. Cells were grown as monolayers and were maintained in exponential growth in 5% CO2 + 95% air in a humidified incubator at 37°C. All cell cultures were confirmed as Mycoplasma free using the Mycoalert® Mycoplasma detection kit (LT07–218, Lonza).
4
Non-Small Cell Lung Cancer Cell Lines
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Human NSCLC cell lines including NCI‐H358, NCI‐H1650, A549, HCC‐827, and NCI‐H1299 were obtained from Cell Bank of the Chinese Academy of Sciences (Chinese Academy of Sciences). Normal human lung epithelial cell line BEAS‐2B was purchased from American Type Culture Collection (ATCC). NCI‐H358, NCI‐H1650, HCC‐827, NCI‐H1299, and BEAS‐2B cell lines were cultured in 90% Roswell Park Memorial Institute (RPMI) 1640 Medium (Gibco) with 10% fetal bovine serum (FBS) (Gibco). A549 cell line was cultured in 90% Ham's F‐12K (Kaighn's) Medium with 10% FBS (Gibco). All cells were cultured in a humidified atmosphere of 95% air and 5% CO2 at 37˚C.
5
Cell Line Cultivation for NSCLC Research
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The human NSCLC cell lines NCI-H1650, PC9, NCI-H1299, NCI-H827, NCI-H520, A549, NCI-H1975, PC14, NCI-H466, NCI-H2170 and NCI-H460 and human umbilical vein endothelial cells (HUVECs) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The human NSCLC cell lines NCI-H1650, NCI-H1299, NCI-H827, NCI-H520, A549, NCI-H1975, PC14, NCI-H466, NCI-H2170 and NCI-H460 were maintained in 1640 medium supplemented with 10% fetal bovine serum (FBS, Gibco) and 1% penicillin/streptomycin (Gibco). The PC9 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS and 1% penicillin/streptomycin (Gibco). The HUVECs were incubated with Ham’s F-12 K supplemented with 100 μg/ml heparin (Sigma), 50 μg/ml endothelial cell growth supplement (BD Biosciences), 10% FBS (Gibco) and 1% penicillin/streptomycin (Gibco).
6
Culturing Diverse Lung Cell Lines
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Human lung bronchial epithelial cell line (BEAS-2B) and human non-small cell lung cancer cell lines (A549, NCI-H1299, NCI-H1650) were all available from ATCC (Manassas, VA, USA) and maintained at 37 °C in an incubator supplied with 5% CO2. Human non-small cell lung cancer cell line PC-9 was purchased from COBIOER Company (Nanjing, China). BEAS-2B cell line was cultivated in BEGM medium (Gibco) with LHC-9 media (Gibco, USA). F-12 K medium (Gibco) was utilized to cultivate A549 cells with 10% FBS. NCI-H1299, NCI-H1650 and PC-9 cells were routinely cultured with 10% FBS in RPMI-1640 medium (Gibco).
7
Culturing Cancer Cell Lines
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The human pancreatic cancer cell line PANC-1, breast cancer cell lines, T-47D and MCF-7, and lung cancer cell lines, NCI-H292 and NCI-H1650, were purchased from the American Type Culture Collection (ATCC, VA, USA). PANC-1 cells were maintained in Dulbecco's minimum essential medium (DMEM; Gibco, NY, USA), while T-47D, MCF-7, NCI-H292 and NCI-H1650 cells were maintained in RPMI-1640 medium (Gibco), supplemented with 10% fetal calf serum (FCS; Hyclone, UT, USA), 100 units/ml penicillin, and 100 mg/ml streptomycin. The cultures were incubated at 37°C in a humidified atmosphere with 5% CO2. Cells were passaged every 2–3 days to obtain exponential growth.
8
Lung Cancer Cell Culture and Clinical Samples
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Non-small cell lung cancer cells A549, NCI-H1650 and PC-9 were purchased from ATCC (USA). A549, NCI-H1650 and PC-9 cells were cultured in RPMI 1640 medium while HEK-293T cell was cultured in DMEM with 10% fetal bovine serum (FBS, Gibco, USA) in condition of 5% CO2 and 37°C. NSCLC and adjacent normal lung tissues were obtained from 50 patients who were diagnosed as NSCLC and treated at The Tumor Hospital of Jilin Province. Clinical researches has been approved by ethics committees.
9
Luciferase-expressing Lung Adenocarcinoma Cells
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Lung adenocarcinoma NCI-H1650 (ATCC, Manassas, VA) and NCI-H1650.LMC cells (NCI-H1650 cells transduced with luciferase and mCherry) were cultured in RPMI 1640 cell culture medium (Gibco, Gaithersburg, MD) supplemented with 10% fetal bovine serum (FBS, Fisher Scientific, Waltham, MA) and 1% penicillin/streptomycin (P/S, Fisher Scientific, Waltham, MA). To generate the NCI-H1650.LMC line, a fusion construct of luc2 (Promega, Madison, WI) and mCherry (Clontech, Mountain View, CA) was cloned into the Lenti-X lentiviral vector (Clontech). NCI-H1650 cells were transduced with lentiviral particles for 48 h and a pool of cells stably expressing the fusion construct were selected using 2 µg/ml puromycin for 2 weeks. All cultures were maintained in a 37 °C humidified incubator with 5% CO2. In all experiments, cells with less than 10 passages after thawing were used.
10
NY-ESO-1 Peptide Presentation on HLA-A2
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The antigen NY-ESO-1 157-165 and HLA-A*02:01 double positive cells were the human tumor lines A375, Mel624, and U266-B1. The antigen NY-ESO-1 157-165 -negative and HLA-A2 positive cells were the melanoma line Mel526 and lung carcinoma line NCI-H1650. A375, U266-B1, and NCI-H1650 were purchased from the American Type Culture Collection. Mel624 and Mel526 were gifts from Prof. Cassin Yee's laboratory. In addition, we also obtained human erythroleukemic cells (K562) transfected with HLA*A02:01 and NY-ESO-1 from Xiangxue Life Sciences Ltd. All tumor cell lines were checked with HLA typing (BFR Gene Diagnostics, Beijing, China) and mRNA expression (Nanostring, Guangzhou, China). The Mel624, U266-B1, Mel526, and NCI-H1650 cells were maintained in RPMI 1640 (GIBCO Life Technologies, Shanghai, China) supplemented with 10% fetal bovine serum (FBS) (GIBCO Life Technologies). The A375 was maintained in Dulbecco's modified Eagle medium (DMEM, GIBCO Life Technologies) supplemented with 10% FBS. The T2 cell line (human lymphoblast) was maintained in RPMI 1640 culture medium supplemented with 10% FBS. The 293T cell line is a human embryonic kidney cell, which was cultured in DMEM supplemented as described previously.
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