Nci h187

Manufactured by American Type Culture Collection

Sourced in United States

The NCI-H187 is a cell line derived from a small cell lung carcinoma. It serves as a model system for research and scientific investigations.

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7 protocols using nci h187

SCLC Cell Lines and Assays

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KP1 (Rb1/Trp53 mutant, DKO), and KP11 (Rb1/Rbl2/Trp53 mutant, TKO) mouse SCLC cells were generated from single tumors (Park et al., 2011 (link)). Allografts tested in the multiplexed inhibitor beads inhibitors assays were derived from single tumors. NJH29 cells were previously generated in our laboratory from a PDX model (Jahchan et al., 2013 (link)), and A549, NCI-H69, NCI-H82, NCI-H187, and NCI-H446 were purchased from ATCC. Cells were grown in RPM1-1640 media supplemented with 10% bovine growth serum (BGS, Fisher Scientific) and penicillin-streptomycin–glutamine (PSQ, Gibco). Cells were grown at 37C in standard cell culture incubators. All cell lines are routinely tested (Lonza) and confirmed to be free of mycoplasma contamination.

Coles G.L., Cristea S., Webber J.T., Levin R.S., Moss S.M., He A., Sangodkar J., Hwang Y.C., Arand J., Drainas A.P., Mooney N.A., Demeter J., Spradlin J.N., Mauch B., Le V., Shue Y.T., Ko J.H., Lee M.C., Kong C., Nomura D.K., Ohlmeyer M., Swaney D.L., Jackson P.K., Narla G., Gordan J.D., Shokat K, & Sage J. (2020). UNBIASED PROTEOMIC PROFILING UNCOVERS A TARGETABLE GNAS/PKA/PP2A AXIS IN SMALL CELL LUNG CANCER STEM CELLS. Cancer cell, 38(1), 129-143.e7.

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Transient Notch Overexpression in SCLC

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Mouse (KP1) and human (NJH29, NCI-H82 and NCI-H187) SCLC cell lines were grown in RPMI-1640 media supplemented with 10% bovine growth serum (BGS) (Fisher Scientific) and penicillin-streptomycin-glutamine (Gibco), as described before⁵⁰. KP1, NJH29 were generated at Stanford. NCI-H82 and NCI-H187 were purchased from ATCC. These cells grow as suspension spheres or aggregates in culture. All cell lines were maintained at 37 °C in a humidified chamber with 5% CO₂. All cell lines tested negative for mycoplasma infection. For transient expression of Notch ICD, cells were trypsinized and transfected with either MigR1-IRES-GFP (Ctrl) or MigR1-Notch1-ICD-IRES-GFP (NICD) using Lipofectamine 2000 (Life Technologies). The plasmids were gifts from W.S. Pear (University of Pennsylvania, Philadelphia). Then 48 h after transfection, cells were trypsinized and resuspended in phosphate-buffered saline (PBS) containing 10% BGS and 1 µg ml⁻¹ 7-aminoactinomycin D (Life Technologies) that labels dead cells. Live GFP⁺ cells were then sorted for subsequent experiments using a BD FACSAria fluorescence-activated cell sorting (FACS) machine.

George J., Lim J.S., Jang S.J., Cun Y., Ozretić L., Kong G., Leenders F., Lu X., Fernández-Cuesta L., Bosco G., Müller C., Dahmen I., Jahchan N.S., Park K.S., Yang D., Karnezis A.N., Vaka D., Torres A., Wang M.S., Korbel J.O., Menon R., Chun S.M., Kim D., Wilkerson M., Hayes N., Engelmann D., Pützer B., Bos M., Michels S., Vlasic I., Seidel D., Pinther B., Schaub P., Becker C., Altmüller J., Yokota J., Kohno T., Iwakawa R., Tsuta K., Noguchi M., Muley T., Hoffmann H., Schnabel P.A., Petersen I., Chen Y., Soltermann A., Tischler V., Choi C.M., Kim Y.H., Massion P.P., Zou Y., Jovanovic D., Kontic M., Wright G.M., Russell P.A., Solomon B., Koch I., Lindner M., Muscarella L.A., la Torre A., Field J.K., Jakopovic M., Knezevic J., Castaños-Vélez E., Roz L., Pastorino U., Brustugun O.T., Lund-Iversen M., Thunnissen E., Köhler J., Schuler M., Botling J., Sandelin M., Sanchez-Cespedes M., Salvesen H.B., Achter V., Lang U., Bogus M., Schneider P.M., Zander T., Ansén S., Hallek M., Wolf J., Vingron M., Yatabe Y., Travis W.D., Nürnberg P., Reinhardt C., Perner S., Heukamp L., Büttner R., Haas S.A., Brambilla E., Peifer M., Sage J, & Thomas R.K. (2015). Comprehensive genomic profiles of small cell lung cancer. Nature, 524(7563), 47-53.

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SCLC and NSCLC Cell Lines Cultivation

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A panel of 9 SCLC cell lines (NCI‐H69, DMS79, NCI‐H187, NCI‐H209, NCI‐H446, NCI‐H510A, NCI‐H526, NCI‐H841, and SW1271">SW1271) and 1 NSCLC cell line (A549) were purchased from ATCC (Manassas, VA, USA). All cell lines were maintained on RPMI‐1640 medium (Gibco, Life Technologies, Carlsbad, CA, USA) supplemented with 10% FBS (Gibco) in a humidified atmosphere of 5% CO₂ at 37°C.

Xu S., Lam S., Cheng P.N, & Ho J.C. (2018). Recombinant human arginase induces apoptosis through oxidative stress and cell cycle arrest in small cell lung cancer. Cancer Science, 109(11), 3471-3482.

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Comprehensive Antimicrobial and Cytotoxicity Assays

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Growth inhibitory activity against M. tuberculosis H₃₇Ra and non-cancerous Vero cells (African green monkey kidney fibroblast, ATCC CCL-81) was evaluated using the green fluorescent protein microplate assay (GFPMA),^{36,37 (link)} The resazurin microplate assay (REMA)^{38 (link)} was used to evaluate cytotoxicity against cancerous cells, including MCF-7 (human breast cancer, ATCC HTC-22) and NCI-H187 (human small-cell lung cancer, ATCC CRL-5804). Antibacterial activity against S. aureus (ATCC 29213) and A. baumannii (ATCC 19606) was evaluated by using standard protocols published by the Clinical and Laboratory Standard Institute.^39,40 5(6)-Carboxyfluorescein diacetate (CFDA)^41–43 fluorometric assay was used to evaluate anti-phytopathogenic fungal activity against C. acutatum (BCC 58146), A. brassicicola (BCC 42724) and C. lunata DOAC 1479 (BCC 15558).

Kornsakulkarn J., Auncharoen P., Khonsanit A., Boonyuen N, & Thongpanchang C. (2023). Cytotoxic cytochalasans from cultures of the fungus Metarhizium brunneum TBRC-BCC 79240. RSC Advances, 13(16), 10564-10576.

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Culturing Various Human and Mouse Cancer Cell Lines

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Human cell lines DMS-273, NCI-H446, NCI-H1304, NCI-H69, NCI-H82, NCI-H187, SW1573, A549, NCI-H226, NCI-H522, SK-MES-1 and H1299 were obtained from the American Type Culture Collection. BJ primary fibroblasts were a kind gift from prof. Reuven Agami, primary MEFs were isolated as previously described (13 (link)). All these cell lines were cultured in DMEM/F12 supplemented with 10% fetal calf serum and 100 units/ml penicillin and 100 μg/ml streptomycin or in RPMI supplemented with 10% fetal calf serum, 1 mM sodium pyruvate and 100 units/ml penicillin and 100 μg/ml streptomycin (all Gibco). LMYC5 mouse SCLC cells were previously generated from a TP53-/- RB1-/- LMYC mouse model (14 ). These cells were grown in DMEM/F12 supplemented with 10% fetal calf serum, HITES (Gibco), and penicillin/streptomycin. All cells were grown in a humidified atmosphere of 5% CO₂ at 37°C.

Nagel R., Avelar A.T., Aben N., Proost N., van de Ven M., van der Vliet J., Cozijnsen M., de Vries H., Wessels L.F, & Berns A. (2019). Inhibition of the replication stress response is a synthetic vulnerability in SCLC that acts synergistically in combination with cisplatin. Molecular cancer therapeutics, 18(4), 762-770.

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Profiling CD63-BCAR4 Fusion in Lung Cancer

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The presence of CD63–BCAR4 fusion transcript and expression of BCAR4 were examined in 24 cell lines. Lung cancer cell lines were purchased from Korean cell line bank (KCLB, Seoul, Korea) (NCI-H187, NCI-H417, NCI-H1299, NCI-H596, NCI-H146, SNU-1327, Hcc1195, Hcc1171, NCI-H1703, NCI-H69, Hcc95, Hcc2108, Hcc2279, A427, A549, NCI-H23, NCI-H460 and NCI-H358) and American type culture collection (ATCC, Manassas, VA, USA) (BEAS-2B, NCI-H226, NCI-H2171, NCI-H1395, NCI-H1975 and NCI-H2228). Immortalised human bronchial epithelial cell line BEAS-2B and breast cancer cell line MDA-MB-231 were purchased from ATCC. Each cell line was cultured in the appropriate medium with 10% foetal bovine serum and 1% penicillin–streptomycin or airway epithelial cell basal medium with bronchial epithelial cell growth kit. The authentication of the cell lines was assessed by using short tandem repeat analysis and mycoplasma infection was tested by PCR. Coding regions of the CD63–BCAR4 fusion and BCAR4 (HE601934.1) were cloned into a lentiviral vector with hemagglutinin (HA) tag, CMV promoter and green fluorescent protein (GFP) as a transduction marker. To establish stable cell lines for CD63–BCAR4, BCAR4 and empty vector, the infected cells were sorted by GFP signal intensity.

Bae K., Kim J.H., Jung H., Kong S.Y., Kim Y.H., Kim S., Lee G.K., Lee J.S., Lee J.J., Ju Y.S., Choi Y.K, & Yoon K.A. (2020). A fusion of CD63–BCAR4 identified in lung adenocarcinoma promotes tumorigenicity and metastasis. British Journal of Cancer, 124(1), 290-298.

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SCLC Cell Line Maintenance Protocol

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All SCLC cell lines, including NCI-H82, NCI-H446, NCI-H69, NCI-H211, NCI-H524, NCI-H526, NCI-H146, NCI-H187, NCI-H345, NCI-H841, NCI-H1930, NCI-H2171, and NCI-H2081, were obtained from American Type Culture Collection (ATCC). NCI-H345, NCI-H841, NCI-H2171, and NCI-H2081 cell lines were maintained in HITES medium [Dulbecco’s medium: Ham’s F12, 50:50 mix (Gibco), insulin 0.005 mg/ml, transferrin 0.01 mg/ml, sodium selenite 30 nM, hydrocortisone 10 nM, and beta-estradiol 10 nM] containing 5% fetal bovine serum (FBS, Hyclone) and penicillin–streptomycin antibiotics (Gibco). The remainder of the cell lines were cultured in RPMI1640 medium (Gibco) supplemented with 10% FBS and antibiotics. Cells were cultured for up to 8 weeks and used at passages of less than 16 from the initial source vial.

Hung M.C., Wang W.P, & Chi Y.H. (2022). AKT phosphorylation as a predictive biomarker for PI3K/mTOR dual inhibition-induced proteolytic cleavage of mTOR companion proteins in small cell lung cancer. Cell & Bioscience, 12, 122.

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