Nci h520

Manufactured by Thermo Fisher Scientific

Sourced in United States

The NCI-H520 is a non-small cell lung cancer cell line derived from a human lung squamous cell carcinoma. It is commonly used in cancer research applications.

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13 protocols using nci h520

Cell Culture Methods for FGFR1-Positive and Negative Lung Cancer

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CHO-S cells (Thermo Fisher Scientific) were cultured in serum-free Power-CHO medium (Lonza) supplemented with 8 mM L-glutamine (Thermo Fisher Scientific) and 1% penicillin and streptomycin mix (Biowest). The cells were subcultered every 2–3 days at a seeding density of 0.2–0.3 × 10⁶ cells·mL⁻¹. The cells were grown at 37°C with 8% CO₂ in a shaking incubator (140 rpm).
NCI-H520 (lung squamous cell carcinoma, FGFR1-positive), NCI-H1581 (lung large cell carcinoma, FGFR1-positive), and HCC95 (lung squamous cell carcinoma, FGFR1-negative) were obtained from the American Type Culture Collection (ATCC). NCI-H520 and NCI-H1581 were cultured in RPMI 1640 (Gibco) with 10% fetal bovine serum (Biowest) and 1% penicillin and streptomycin mix (Biowest); HCC95 cells were cultured in RPMI 1640 (Gibco) with 10% fetal bovine serum (Biowest), sodium bicarbonate (Gibco), and 1% penicillin and streptomycin mix (Biowest). The cancer cell lines were cultured at 37°C with 5% CO₂.

Jendryczko K., Rzeszotko J., Krzyscik M.A., Szymczyk J., Otlewski J, & Szlachcic A. (2021). Peptibody Based on FGFR1-Binding Peptides From the FGF4 Sequence as a Cancer-Targeting Agent. Frontiers in Pharmacology, 12, 748936.

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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).

Wang T., Xing Y., Meng Q., Lu H., Liu W., Yan S., Song Y., Xu X., Huang J., Cui Y., Jia D, & Cai L. (2019). Mammalian Eps15 homology domain 1 potentiates angiogenesis of non-small cell lung cancer by regulating β2AR signaling. Journal of Experimental & Clinical Cancer Research : CR, 38, 174.

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BSO Treatment of NSCLC Cell Lines

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NCI-H520 and SK-MES-1 were purchased from Cellcook Co., Ltd. (Guangzhou, China) with cell authentication via the STR multi-amplification method. NCI-H520 was cultured in RPMI1640 (Gibco BRL). SK-MES-1 was cultured in Eagle’s Minimum Essential Medium. All medium contained 10% FBS and 1% penicillin/streptomycin. The cells were cultured at 37°, under 5% CO₂. BSO (yuanye, Shanghai, China) was dissolved in H₂O. BSO was diluted to 249.7uM, followed by replacing the cell medium 12h after cells seeded. All experiments were completed less than 2 months after establishing stable cell lines or thawing early passage cells.

Zhang X., Zhang R., Liu P., Zhang R., Ning J., Ye Y., Yu W, & Yu J. (2022). ATP8B1 Knockdown Activated the Choline Metabolism Pathway and Induced High-Level Intracellular REDOX Homeostasis in Lung Squamous Cell Carcinoma. Cancers, 14(3), 835.

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Lung Squamous Cell Carcinoma Cell Lines

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The human lung SCC cell lines NCI-H520, NCI-H226 and SK-MES-1, as well as the human normal lung cell line BEAS-2B, were all purchased from the American Type Culture Collection (VA, U.S.A.); the human lung SCC cell line LTEP-s was purchased from BeNa Culture Collection (Beijing, China). For cell culture, LTEP-s cells were cultured in D MEM with high glucose (Gibco, CA, U.S.A.); NCI-H520 and NCI-H226 cells were cultured in RPMI-1640 medium (Gibco, CA, U.S.A.); SK-MES-1 cells were cultured in MEM (Gibco, CA, U.S.A.), all with 10% foetal bovine serum (FBS; Gibco, CA, U.S.A.) and 1% penicillin and streptomycin supplement. All cells were maintained in a humidified incubator at 37°C with 5% CO₂.

Wang D., Gao Y., Zhang Y., Wang L, & Chen G. (2019). Glypican-3 promotes cell proliferation and tumorigenesis through up-regulation of β-catenin expression in lung squamous cell carcinoma. Bioscience Reports, 39(6), BSR20181147.

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Characterization of NSCLC Cell Lines

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Non-cancerous, immortalized human lung bronchial epithelial cell line BEAS-2B was purchased from Zhong Qiao Xin Zhou Biotechnology (Shanghai, China). Normal fetal lung fibroblast cell HFL1; human LUAD cell lines Calu-3, NCI-H1975, and NCI-H1395; LUSC line NCI-H520; and other NSCLC cell lines A549 and NCI-H1299 were purchased from Procell (Wuhan, China). LUSC lines NCI-H226 and SK-MES-1 were purchased from Shanghai Chinese Academy of Science Cell Bank (Shanghai, China). BEAS-2B was grown in Bronchial Epithelial Cell Growth Medium BulletKit (BEGM; Lonza, Basel, Switzerland). HFL1 was cultured in Ham’s F-12K medium (Gibco, Pittsburgh, PA, USA). Calu-3 and SK-MES-1 were cultured in Eagle’s minimum essential medium (MEM; Gibco). NCI-H1975, NCI-H1395, NCI-H1299, NCI-H520, and NCI-H226 were maintained in RPMI-1640 (Gibco). All cell lines, except for BEAS-2B, were supplemented with 10% fetal bovine serum (FBS), streptomycin (100 μg/mL), and penicillin G (100 U/mL; Gibco). All cell lines were authenticated by short tandem repeat (STR) profiling. Cells were incubated at 37°C in a humidified atmosphere with 5% CO₂.

Liu A., Xie H., Li R., Ren L., Yang B., Dai L., Lu W., Liu B., Ren D., Zhang X., Chen Q., Huang Y, & Shi K. (2021). Silencing ZIC2 abrogates tumorigenesis and anoikis resistance of non-small cell lung cancer cells by inhibiting Src/FAK signaling. Molecular Therapy Oncolytics, 22, 195-208.

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Cell Culture Protocols for Cancer Research

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DMS114 (human
small cell lung cancer, ATCC CRL-2066)
cells were cultured in Waymouth’s MB 752/1 from Gibco (Waltham,
MA). U2OS (human osteosarcoma, HTB-96) and U2OS stably transfected
with FGFR1 (U2OS-R1) were cultured in DMEM High Glucose with stable
glutamine and sodium pyruvate from Biowest (France). NCI-H520 (human
squamous cell carcinoma, ATCC HTB-182) and HCC15 (human squamous cell
lung carcinoma) and were cultured in RPMI-1640 medium from Gibco (Waltham,
MA). All media were supplemented with 10% fetal bovine serum from
Thermo Fisher Scientific, and 1% penicillin/streptomycin mix was from
Biowest (France). Additionally, the U2OS-R1 cell medium contained
50 μg/mL gentamicin sulfate from Thermo Fisher Scientific. All
cell lines were cultured in a humidified incubator at 37 °C in
a 5% CO₂ atmosphere. The DMS114, NCI-H520, and U2OS cell
lines were obtained from American Type Culture Collection (Manassas,
VA). HCC15 cells were supplied by the Leibniz Institute DSMZ, German
Collection of Microorganisms and Cell Cultures. The U2OS cells stably
expressing FGFR1 (U2OS-R1) were a kind gift from Dr. Ellen M. Haugsten
from The Norwegian Radium Hospital.^{38 (link)} The E. coli expression strain Rosetta 2(DE3)pLysS was from Novagen-EMD
Biosciences (Madison, WI).

Krzyscik M.A., Zakrzewska M, & Otlewski J. (2020). Site-Specific, Stoichiometric-Controlled, PEGylated Conjugates of Fibroblast Growth Factor 2 (FGF2) with Hydrophilic Auristatin Y for Highly Selective Killing of Cancer Cells Overproducing Fibroblast Growth Factor Receptor 1 (FGFR1). Molecular Pharmaceutics, 17(7), 2734-2748.

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Cell Culture and Lentiviral Transduction

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ASPC1, HGC27, MKN45, NCI-H520, Hut78 and Jurkat cells were purchased from Nanjing Cobioer Biosciences (Nanjing, China). The cells were tested via short tandem repeat (STR) profiling. ASPC1, HGC27, MKN45, NCI-H520 and Jurkat cells were cultured in RPMI1640 (22400-089, Gibco, USA) containing 10% fetal bovine serum (10091-148, Gibco, USA). ASPC1-luc, HGC27-luc, MKN45-luc, and NCI-H520-luc were constructed by lentivirus infection (H7656, OBiO Technology, China) and cultured in RPMI1640 containing 10% fetal bovine serum, and 2 μg/ml Puromycin (A11138-02, Gibco, USA). Hut78 was cultured in IMDM containing 10% fetal bovine serum. All cell lines were cultured in a humidified incubator with 5% CO2 at 37 °C.

Ye J., Liu Q., He Y., Song Z., Lin B., Hu Z., Hu J., Ning Y., Cai C, & Li Y. (2024). Combined therapy of CAR-IL-15/IL-15Rα-T cells and GLIPR1 knockdown in cancer cells enhanced anti-tumor effect against gastric cancer. Journal of Translational Medicine, 22, 171.

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Cell Culture Conditions for Diverse Cell Lines

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CHO-K1SV, NCI-H520, A549, H358 and MRC-5 cells were purchased from American Type Culture Collection (Manassas, VA, USA). NCI-H520, A549 and H358 cells were cultured in Dulbecco's modified Eagle's medium, supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA). MRC-5 cells were cultured in RPMI-1640 supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.). All cells were cultured at 37°C in an environment containing 5% CO₂.

Shi W., Song J., Wang W., Zhang Y, & Zheng S. (2017). MACC-1 antibody target therapy suppresses growth and migration of non-small cell lung cancer. Molecular Medicine Reports, 16(5), 7329-7336.

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Lung Cancer Cell Line Characterization

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NCI-H520 and SK-MES-1 were purchased from Cellcook Co., Ltd. with cell authentication via STR multi-amplification method. KLN205 was obtained from Chinese Academy of Medical Sciences tumor cell libraries. NCI-H520 was cultured in RPMI1640 (Gibco BRL). SK-MES-1 was cultured in MEM. KLN205 was cultured in H-DMEM. All medium contained 10% FBS and 1% penicillin/streptomycin. For 12/15-LOX inhibitor treatment experiments, ML355 and PD146176 was diluted in medium, followed by replacing the cell medium 5 h after cells seeded respectively. Cell lines were routinely evaluated for Mycoplasma contamination. All experiments were completed less than 2 months after establishing stable cell lines or thawing early-passage cells.

Sun Z., Zhang R., Zhang X., Sun Y., Liu P., Francoeur N., Han L., Lam W.Y., Yi Z., Sebra R., Walsh M., Yu J, & Zhang W. (2022). LINE-1 promotes tumorigenicity and exacerbates tumor progression via stimulating metabolism reprogramming in non-small cell lung cancer. Molecular Cancer, 21, 147.

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Lung Cancer Cell Line Cultivation

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Two human lung cancer cell lines NCI-H520 (squamous cell carcinoma) and NCI-H23 (adenocarcinoma) as well as normal cell line MRC-9 (human lung fibroblast cells) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Both NCI-H520 and NCI-H23 cell lines were grown in RPMI 1640 media while the MRC-9 cell line was grown in Eagle’s Minimum Essential Medium (EMEM, Gibco, Grand Island, NY, USA). All of the cell lines were maintained in respective media supplemented with 10% fetal bovine serum (FBS, Gibco) and 1% penicillin (100 U/mL)/streptomycin (100 µg/mL) (Gibco) in a humidified atmosphere with 5% CO₂ at 37 °C. For sustaining the growth of the cells, the medium in the flask was changed at every two–three-day interval until 80–90% of growth confluency was achieved. Upon achieving 90% confluency, the cells were subcultured using accutase (Gibco) as the cell detachment solution. All of the cell culture procedures were performed in a biosafety cabinet and appropriate aseptic techniques were adhered strictly to prevent contamination.

Wan Mohd Tajuddin W.N., Abas F., Othman I, & Naidu R. (2021). Molecular Mechanisms of Antiproliferative and Apoptosis Activity by 1,5-Bis(4-Hydroxy-3-Methoxyphenyl)1,4-Pentadiene-3-one (MS13) on Human Non-Small Cell Lung Cancer Cells. International Journal of Molecular Sciences, 22(14), 7424.

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