Calu 6

Manufactured by American Type Culture Collection

Sourced in United States

The Calu-6 is a cell line derived from a human lung carcinoma. It is a commonly used in vitro model for studying various aspects of lung cancer biology. The Calu-6 cell line maintains key characteristics of the original tumor and is a valuable tool for cancer research.

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Calu-6 cells (5 citations) SCLC Calu-6 cells (2 citations)

75 protocols using calu 6

NSCLC Cell Line Maintenance and Chemical Treatment

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NSCLC cell lines H1299, Calu-6, SK-MES-1, H2006, H460 and H1437 were purchased from ATCC and maintained in DMEM (H1299); MEM (Calu-6 and SK-MES-1) or RPMI1640 (H2006, H460 and H1437) medium supplemented with 10% FBS and 1% penicillin/streptomycin. Cell lines were routinely screened for mycoplasma contamination using LookOut^® Mycoplasma PCR detection kit from Sigma (Cat. MP0035). Cells were cultured at 37°C in a humidified incubator at 95% humidified oxygen and 5% CO2. The CHK1 inhibitor LY2603618 was purchased from APExBIO cat. #A8638, and the ATR inhibitor VE-821 (cat. #S8007) was purchased from Selleckchem. Auranofin (Cat. #A6733) and N-Acetyl cysteine (cat. #A9165) and Iodoacetamide (IAA, cat. #A3221) were purchased from Sigma Aldrich.

Prasad C.B., Oo A., Qiu Z., Li N., Singh D., Xin X., Cho Y.J., Li Z., Zhang X., Yan C., Zheng Q., Shao J., Wang Q.E., Kim B, & Zhang J. (2023). The thioredoxin system determines CHK1 inhibitor sensitivity via redox-mediated regulation of ribonucleotide reductase activity. Research Square.

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Targeted Lung Cancer Treatment Efficacy

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Example 13

[Figure (not displayed)]

Human non-small cell lung cancer line Calu-6 cells were purchased from ATCC (American Type Culture Collection). 5×10⁶cells suspended in physiological saline were subcutaneously transplanted to the right abdomen of each female nude mouse (Day 0), and the mice were randomly grouped at Day 11. The M30-H1-L4P antibody and antibody-drug conjugate (2) were each intravenously administered at a dose of 10 mg/kg to the tail of each mouse at Days 11, 18, and 25 in a schedule of qw×3.

The results are shown in FIG. 19. In the drawing, the line with open rhombuses depicts the results about untreated tumor, the line with open triangles depicts the effect of the M30-H1-L4P antibody, and the line with open circles depicts the effect of the antibody-drug conjugate (2). The administration of the antibody-drug conjugate (2) remarkably decreased the tumor volume, and no further tumor growth was observed after the final administration. By contrast, the administration of the M30-H1-L4P antibody resulted in the progression of tumor growth.

In addition, the mice that received the antibody-drug conjugate (2) were free from notable signs such as weight loss, suggesting that the antibody-drug conjugate (2) is low toxic and highly safe.

US10973924B2. Antibody-drug conjugate (2021-04-13). Daiichi Sankyo Company, Limited [JP]. Inventors: Takeshi Masuda [JP], Hiroyuki Naito [JP], Takashi Nakada [JP], Masao Yoshida [JP], Shinji Ashida [JP], Hideki Miyazaki [JP], Yuji Kasuya [JP], Koji Morita [JP], Yuki Abe [JP], Yusuke Ogitani [JP].

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Cell Line Provenance and Sterility

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COS7, HEK293T, K562, HeLa, Calu-6 were purchased from ATCC and early-passage stocks were in order to ensure cell line provenance and sterility. The BGC-823 cell line was purchased from the Istituto Nazionale per la Ricerca sul Cancro (Genova, Italy) and early-passage stocks were used in order to ensure cell line provenance and sterility.

D’Lima N.G., Ma J., Winkler L., Chu Q., Loh K.H., Corpuz E.O., Budnik B.A., Lykke-Andersen J., Saghatelian A, & Slavoff S.A. (2016). A human microprotein that interacts with the mRNA decapping complex. Nature chemical biology, 13(2), 174-180.

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Cancer Cell Culture Protocol

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MCF-7 (ATCC: HTB-22, breast cancer cell line), DU145 (ATCC: HTB-81, prostate cancer cell line), Calu-6 (ATCC: HTB-56, lung cancer cell line), A498 (ATCC: HTB-44, kidney cancer cell line), SK-OV-3 (ATCC: HTB-77, ovarian cancer cell line), HT-29 (ATCC: HTB-38, colon cancer cell line), PC-3 (ATCC: CRL-1435), OVCAR-3 (ATCC: HTB-161, ovarian cancer cell line), A549 (ATCC: CCL-185, lung cancer cell line), and MDA-MB-231 (ATCC: HTB-26, breast cancer cell line) were grown at 37 °C and 5% CO₂ in minimal essential medium Eagle (MEM) cell culture medium supplemented with 10% fetal bovine serum and, 1% L-glutamine. Trypsin-EDTA, minimal essential medium Eagle (MEM), L-glutamine, fetal bovine serum and cell culture flasks (75 cm², surface treated with filter cap) were purchased from PAA Laboratories (Somerset, UK). Hanks balanced salt solution was obtained from Sigma–Aldrich (Dorset, UK).

Khan H., Makwana V., dos Santos S.N., Bonacossa de Almeida C.E., Santos-Oliveira R, & Missailidis S. (2021). Development, Characterization, and In Vivo Evaluation of a Novel Aptamer (Anti-MUC1/Y) for Breast Cancer Therapy. Pharmaceutics, 13(8), 1239.

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Cell Line Authentication and Characterization

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Cell lines were cultured according to the provider’s instruction and authenticated by STR analysis. A101D, A549, AsPC-1, BxPC-3, Caki-1, Calu-6, Hs 746 T, LoVo, MIA PaCa-2, NCI-H2122, PANC-1, PC-3, SK-OV-3, SW480 and TOV-21G were from the ATCC. A2780 and A-498 were from the ECACC and DSMZ, respectively. HCC-461 was a kind gift from Dr. Heidi Greulich (Dana Farber Cancer Institute).

Hirt U.A., Waizenegger I.C., Schweifer N., Haslinger C., Gerlach D., Braunger J., Weyer-Czernilofsky U., Stadtmüller H., Sapountzis I., Bader G., Zoephel A., Bister B., Baum A., Quant J., Kraut N., Garin-Chesa P, & Adolf G.R. (2018). Efficacy of the highly selective focal adhesion kinase inhibitor BI 853520 in adenocarcinoma xenograft models is linked to a mesenchymal tumor phenotype. Oncogenesis, 7(2), 21.

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Cell Line Selection and Authentication

Cited 2 times

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Cell lines were selected from a previously published panel located in the Center for Molecular Therapeutics at Massachusetts General Hospital (30 (link)), except for A549 and Calu-6 which were purchased directly from ATCC, and DLD1 (KRAS G13D/wt) and DWT7 (del/wt) which were kind gifts from Dr. Bert Vogelstein (34 (link)). The identity of each of the cell lines in the panel had been tested as described previously (6 (link)), and additional cell line authentication was performed by Bio-Synthesis, Inc. No cell line was ever treated for mycoplasma and all lines tested mycoplasma free prior to the experiments (MycoAlert, Lonza). NCI-H1703 cells harboring wild-type KRAS were transfected with a pBABE-Puro vector encoding KRAS G12V or an empty control (kindly provided by Dr. David Barbie). Stably transfected clones were selected with 2 mg/mL puromycin (Sigma, P9620) and maintained at 1 mg/mL puromycin. shRNA KRAS transfection of A549 cells was performed as described previously (35 (link)). For 3D culture of tumor spheres, ~5,000 cells/well were grown in low-binding 96-well plates (Thermo, 145399) using serum-free medium composed of DMEM (Sigma-Aldrich), basic fibroblast growth factor (bFGF), and EGF (20 ng/mL each, Sigma-Aldrich), and B27 supplement (Life Technologies).

Wang M., Kern A.M., Hülskötter M., Greninger P., Singh A., Pan Y., Chowdhury D., Krause M., Baumann M., Benes C.H., Efstathiou J.A., Settleman J, & Willers H. (2014). EGFR-Mediated Chromatin Condensation Protects KRAS-Mutant Cancer Cells Against Ionizing Radiation. Cancer research, 74(10), 2825-2834.

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Antibody-Drug Conjugate Inhibits Lung Cancer

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Example 13

[Figure (not displayed)]

In addition, the mice that received the antibody-drug conjugate (2) were free from notable signs such as weight loss, suggesting that the antibody-drug conjugate (2) is low toxic and highly safe.

US10117952B2. Antibody-drug conjugate (2018-11-06). Daiichi Sankyo Company, Limited [JP]. Inventors: Takeshi Masuda [JP], Hiroyuki Naito [JP], Takashi Nakada [JP], Masao Yoshida [JP], Shinji Ashida [JP], Hideki Miyazaki [JP], Yuji Kasuya [JP], Koji Morita [JP], Yuki Abe [JP], Yusuke Ogitani [JP].

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Culturing Human NSCLC Cell Lines

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Human non–small cell lung cancer (NSCLC) cell lines Calu-6 and A549 were purchased from the ATCC. HAP-1 DNAPK wild-type (WT) and knockout cell lines were purchased from Horizon Discovery. All cell lines were Mycoplasma negative as tested by PCR. Cells were cultured in advanced DMEM/F12 medium (GIBCO/Life Technologies), supplemented with 5% FBS, 2 mmol/L glutamax, and 50 μg/mL penicillin/streptomycin in a humidified atmosphere with 7.5% CO₂.

Jiang Y., Willmore E., Wedge S.R, & Ryan A.J. (2021). DNAPK Inhibition Preferentially Compromises the Repair of Radiation-induced DNA Double-strand Breaks in Chronically Hypoxic Tumor Cells in Xenograft Models. Molecular Cancer Therapeutics, 20(9), 1663-1671.

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Cell Line Maintenance and Modifications

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The A549, Calu-6, Colo205, DU-145, HCC1569, HEK293, JIMT-1, MDA-MB-231, MDA-MB-453, NCI-H441, NCI-H1569, NS-1, PC-3, SK-BR-3, SP2/0 and 22RV-1 cell lines used were purchased from ATCC and maintained in the recommended medium. HEK293-ERBB3 cells were purchased from Kyinno Biotechnology (KC-1496). SP2/0-HER3 cells stably expressing human HER3 were generated in house by transduction of human HER3 with lentivirus construct.

Li X., Yao J., Qu C., Luo L., Li B., Zhang Y., Zhu Z., Qiu Y, & Hua H. (2024). DB-1310, an ADC comprised of a novel anti-HER3 antibody conjugated to a DNA topoisomerase I inhibitor, is highly effective for the treatment of HER3-positive solid tumors. Journal of Translational Medicine, 22, 362.

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Statins Overcome Erlotinib Resistance in NSCLC

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Experiments were carried out with human lung adenocarcinoma cell lines A549 (ATCC CCL_185), Calu6 (ATCC HTB-56) and NCI-H1993 (ATCC CRL-5909). All cell lines were obtained from American Type Cell Culture Collection (ATCC) and cultured in DMEM growth medium (GIBCO #31966-21), supplemented with 10% heat-inactivated foetal bovine serum (FBS) and 1% antibiotics (penicillin, streptomycin). Cells were kept at 37 °C and 5% CO₂ in the incubator and were passaged at 80–90% confluence every 2–3 days to maintain continuous logarithmic growth. All cell lines studied in this work were erlotinib resistant and EGFR wild type (Table 1). Cells were treated with pitavastatin calcium (SelleckChem, #S1759), fluvastatin sodium (SelleckChem, #S1909) and erlotinib hydrochloride (SelleckChem, #S1023).Table 1

Human NSCLC cell lines harbouring different genetic mutations examined in the study.

Cell line	EGFR	K-Ras	Met	p53	Erlotinib
A549	Wildtype	Mutated	Wildtype	Wildtype	Resistant
Calu6	Wildtype	Mutated	Wildtype	Mutated	Resistant
H1993	Wildtype	Wildtype	Amplified	Mutated	Resistant

Otahal A., Aydemir D., Tomasich E, & Minichsdorfer C. (2020). Delineation of cell death mechanisms induced by synergistic effects of statins and erlotinib in non-small cell lung cancer cell (NSCLC) lines. Scientific Reports, 10, 959.

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