H1975

Manufactured by Thermo Fisher Scientific

Sourced in United States, China, Germany

The H1975 is a laboratory equipment product manufactured by Thermo Fisher Scientific. It serves as a core function for laboratory tasks, but a detailed description while maintaining an unbiased and factual approach is not available.

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NCI-H1975 (31 citations)

82 protocols using h1975

Culturing and Modulating Lung Cell Lines

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All cell lines were purchased from the American Type Culture Collection (ATCC) (Rockville, MD, USA). Human bronchial epithelial BEAS-2B cells were maintained in the previously described culturing condition [59 (link)]. Human lung cancer cell lines (A549, H1299, H1975, H358, H460, and 95D) were cultured in complete growth medium containing 10% fetal bovine serum (Gibco, New York, NY, USA) and 1% penicillin-streptomycin (Gibco, New York, NY, USA), at 37 degrees Celsius in an atmosphere of 5% CO₂/95% air.
For the establishment of stable CYP27C1-knockdown A549, H1975 cell lines, and stable CYP27C1-overexpressed H460 cell line, plasmids (pLKO.1 TRC-shCYP27C1, pcDNA4-His/Max B-CYP27C1) were transfected into cells, respectively, followed by cell selection using 2 μg/mL of puromycin or 400 μg/mL of zeocin (Thermo Fisher Scientific, Waltham, MA, USA).

Mo H.Y., Wei Q.Y., Zhong Q.H., Zhao X.Y., Guo D., Han J., Noracharttiyapot W., Visser L., van den Berg A., Xu Y.M, & Lau A.T. (2022). Cytochrome P450 27C1 Level Dictates Lung Cancer Tumorigenicity and Sensitivity towards Multiple Anticancer Agents and Its Potential Interplay with the IGF-1R/Akt/p53 Signaling Pathway. International Journal of Molecular Sciences, 23(14), 7853.

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Establishing Lung Cancer Cell Lines for Research

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Lung adenocarcinoma cell lines, A549, SPC‐A1, PC9, H1975 and H1299 and a normal human bronchial epithelial cell line (HBE) were purchased from the Institute of Biochemistry and Cell Biology of the Chinese Academy of Sciences (Shanghai, China). A549, SPC‐A1, H1975 and H1299 were maintained in Roswell Park Memorial Institute (RPMI) 1640 basic medium (GIBCO‐BRL, Invitrogen, Carlsbad, CA) containing 10% foetal bovine serum (FBS), 100 U/l penicillin and 0.1 mg/ml streptomycin. PC9 and HBE cells were maintained in Dulbecco's modified Eagle's medium (DMEM) (GIBCO‐BRL, Invitrogen, Carlsbad, CA, USA) containing 10% foetal bovine serum (FBS), 100 U/l penicillin and 0.1 mg/ml streptomycin. All cells were cultured in a humidified incubator at 37°C with 5% CO₂.

Wang X., Xu W., Zhan P., Xu T., Jin J., Miu Y., Zhou Z., Zhu Q., Wan B., Xi G., Ye L., Liu Y., Gao J., Li H., Lv T, & Song Y. (2018). Overexpression of geranylgeranyl diphosphate synthase contributes to tumour metastasis and correlates with poor prognosis of lung adenocarcinoma. Journal of Cellular and Molecular Medicine, 22(4), 2177-2189.

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Culturing Mouse Lung Carcinoma and Human NSCLC Cells

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Mouse Lewis lung carcinoma (LLC) cell line and human NSCLC cell line (H1975) were purchased from the Cell bank, Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). LLC cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, Beijing, China) containing 10% fetal bovine serum (FBS, ExCell Bio, Shanghai, China) and 1% Penicillin Streptomycin solution (Yu et al., 2020a (link)). H1975 cells were cultured in RPMI 1640 Medium (Invitrogen, 11875-093) with 10% FBS, 1% Glutamax (Invitrogen, 35050061), and 1% Sodium Pyruvate 100 mM Solution (Invitrogen, 11360070) (Jiang et al., 2021 (link)). All cells were cultured in a humidified incubator at 37°C and 5% CO₂.

Shi J., Hao S., Liu X., Li Y, & Zheng X. (2023). Feiyiliu Mixture sensitizes EGFRDel19/T790M/C797S mutant non-small cell lung cancer to osimertinib by attenuating the PRC1/Wnt/EGFR pathway. Frontiers in Pharmacology, 14, 1093017.

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Lung Cancer Cell Line Characterization and RPL6 Knockdown

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The lung cancer cell lines A549, H1975, and H1299 and the normal cell line BEAS-2B were obtained from American Type Culture Collection. All cells were maintained in RPMI-1640 medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) with 10% fetal bovine serum (Gibco, Thermo Fisher Scientific, USA) and 1% penicillin/streptomycin (Sigma, Merck KGaA, USA) in a humidified incubator containing 5% CO₂ at 37 °C. RPL6 knockdown was performed in H1299 and H1975 cells using Lipofectamine RNAiMAX transfection reagent (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA). The siRNAs were purchased from GeneChem, and the sequences were as follows: siRPL6#1: 5'-GCGCAAGAUUGAUCAGAAATT-3'; siRPL6#2: 5'-CUGCCAUGUAUUCCAGAAATT-3'; siCtrl: 5'-UUCUCCGAACGUGUCACGU-3'.

Zhang J., Ma Q., Han Y., Wen H., Zhang Z., Hao Y., Xiao F, & Liang C. (2022). Downregulated RPL6 inhibits lung cancer cell proliferation and migration and promotes cell apoptosis by regulating the AKT signaling pathway. Journal of Thoracic Disease, 14(2), 507-514.

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Lung Adenocarcinoma Cell Line Maintenance and Genetic Manipulation

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Human lung adenocarcinoma cell lines, (H1355, H1573, H23, A549, H1975, and H441) were purchased from the American Type Culture Collection (ATCC) cell bank. The lung adenocarcinoma cell lines CL1-0 and CL1-5 were established and provided as a gift from Dr. Pan-Chyr Yang (National Taiwan University, Taipei, Taiwan). H1355, H1573, H23, CL1-0, H1975, H441, and CL1-5 cells were maintained in RPMI 1640 medium, supplemented with 10% fetal bovine serum (FBS) (Invitrogen, Carlsbad, CA, USA). The human lung adenocarcinoma cell line A549 was grown in F12K medium plus 10% FBS (Invitrogen) at 37 °C in a humidified atmosphere of 5% CO₂. For establishing the stable PGK1 or HTATSF1 cell lines, the pGIPZ lentiviral shRNAmir system (Thermo, Waltham, MA, USA) was used with PGK1 and HTATSF1 sequences, respectively. The lentiviruses were used to infect CL1-5 cells for 2 days. Stable clones were selected with puromycin (Sigma-Aldrich, St. Louis, MO, USA) at 1 μg/ml for 2 weeks.

Chang Y.C., Chan M.H., Li C.H., Yang C.J., Tseng Y.W., Tsai H.F., Chiou J, & Hsiao M. (2021). Metabolic protein phosphoglycerate kinase 1 confers lung cancer migration by directly binding HIV Tat specific factor 1. Cell Death Discovery, 7, 135.

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Characterization of NSCLC Cell Lines

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Human NSCLC cell lines H1975 (adenocarcinoma), H1703 (squamous carcinoma), and A549 (squamous carcinoma) were purchased from American Type Culture Collection (ATCC, Manassas, VA). ATCC characterizes each cell line using cytochrome C oxidase I (COI) and short tandem repeat (STR) testing. H1975 and H1703 were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS, Invitrogen, Carlsbad, CA) and 1.0% penicillin-streptomycin at 37°C in an atmosphere of 5% CO₂. A549 was maintained in Roswell Park Memorial Institute medium (RPMI-1640, Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS, Invitrogen, Carlsbad, CA) and 1.0% penicillin-streptomycin at 37°C in an atmosphere of 5% CO₂.

Dorsey J.F., Kao G.D., MacArthur K.M., Ju M., Steinmetz D., Wileyto E.P., Simone CB I.I, & Hahn S.M. (2014). Tracking Viable Circulating Tumor Cells (CTCs) in the Peripheral Blood of Non-Small Cell Lung Cancer Patients Undergoing Definitive Radiation Therapy: Pilot Study Results. Cancer, 121(1), 139-149.

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Culturing NSCLC and Normal Lung Cells

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Human NSCLC cell lines (A549, PC9, H1299, H1975 and HCC827) and normal pulmonary epithelial cells (BEAS-2B) were obtained from the American Type Culture Collection. A549, PC9, H1299, H1975 and HCC827 cells were cultured in RPMI-1640 medium (Invitrogen; Thermo Fisher Scientific, Inc.). BEAS-2B cells were cultured in DMEM (Invitrogen; Thermo Fisher Scientific, Inc.). All media were supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and all cells were incubated at 37°C with 5% CO_2.

Zhang Y., Liu S., Zhao T, & Dang C. (2021). METTL3-mediated m6A modification of Bcl-2 mRNA promotes non-small cell lung cancer progression. Oncology Reports, 46(2), 163.

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Overexpressing FOXM1 in NSCLC cell lines

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Human NSCLC cell lines, A549, H1299, and H1975 cells were purchased from Shanghai Cell Bank of the Chinese Academy of Sciences. A549 cells were cultured with DMEM medium and H1299 and H1975 cells were grown in RPMI 1640 medium. Both DMEM medium and RPMI 1640 medium were supplemented with 10% fetal bovine serum, penicillin (100 U/ml) and streptomycin sulfate (100 μg/ml). Cells were maintained at 37°C in a humidified 5% CO2 atmosphere. The empty plasmid EX-NEG-M02 (Genecopoeia) and human FOXM1-overexpression plasmid (Genecopoeia) were transfected into A549, H1299, and H1975 cells using Lipofectamine 3,000 reagent (Invitrogen #2024201, MA, United States), according to the manufacturer’s instructions. After that, the A549, H1299, and H1975 cells transfected with the plasmid were used in subsequent experiments.

Ni L., Sun P., Fan X., Li Z., Ren H, & Li J. (2022). Berberine Inhibits FOXM1 Dependent Transcriptional Regulation of POLE2 and Interferes With the Survival of Lung Adenocarcinoma. Frontiers in Pharmacology, 12, 775514.

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NSCLC Cell Line Cultivation and 5-Aza Treatment

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Human NSCLC cell lines H1299, H1975, and A549 were acquired from the American Type Culture Collection (ATCC) (Manassas, VA, USA). H1299 and H1975 cells were cultured in RPMI1640 Medium (Invitrogen, Carlsbad, CA, USA) added with 10% fetal bovine serum (FBS) (Invitrogen). A549 cells were cultured in F12K Medium (Invitrogen) added with 10% FBS. All cells were maintained in a humidified incubator containing 5% CO₂ at 37°C. Where indicated, A549 and H1299 cells were treated with 2.5 μM or 5 μM 5-Aza (Sigma-Aldrich, Merck KGaA, Darmstadt, Germany) for 3 days.

Zhang M., Wu J., Zhong W., Zhao Z, & He W. (2021). DNA-methylation-induced silencing of DIO3OS drives non-small cell lung cancer progression via activating hnRNPK-MYC-CDC25A axis. Molecular Therapy Oncolytics, 23, 205-219.

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Cell Line Cultivation Conditions

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The tumor cell lines A431, A549, H292, HCC827, H1975, and H1755 were obtained from the American Type Culture Collection (ATCC; Lockville, MD). The breast cancer cell line Hs578T was kindly gifted by Dr. Tohru Mochizuki (Shizuoka Cancer Center Research Institute, Japan). A549, H292, HCC827, H1975, and H1755 cells were cultured in RPMI-1640 (Invitrogen, Carlsbad, CA) containing 10% fetal bovine serum (Gibco, Life Technologies, Grand Island, NY). A431 and Hs578T cells was cultured in Dulbecco’s modified Eagle’s medium (DMEM; Invitrogen) containing 10% FBS. Cell lines were cultured under humidified 5% CO₂/95% air at 37°C.

Watanabe M., Serizawa M., Sawada T., Takeda K., Takahashi T., Yamamoto N., Koizumi F, & Koh Y. (2014). A novel flow cytometry-based cell capture platform for the detection, capture and molecular characterization of rare tumor cells in blood. Journal of Translational Medicine, 12, 143.

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