Pc9 cells

Manufactured by Merck Group

Sourced in United States

PC9 cells are a type of lung cancer cell line that are widely used in cancer research. They are derived from human lung adenocarcinoma tissue and exhibit characteristics of non-small cell lung cancer. PC9 cells are commonly utilized in various in vitro studies, including drug screening, cell biology, and molecular biology experiments.

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9 protocols using pc9 cells

Generation of Drug-Tolerant Cancer Cells

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PC9 cells (obtained from Sigma, St. Louis, MO, USA) were cultured in RPMI (Thermo, Waltham, MA, USA) supplemented with 10% FBS (v/v) (Gemini, Dublin, Ireland) and 1% penicillin–streptomycin (Thermo). SKMEL28 cells (obtained from ATCC, Manassas, VA, USA) were cultured in MEM (Thermo) supplemented with 10% FBS (v/v) and 1% penicillin–streptomycin. Drug-naive, early-passage PC9 and SKMEL28 cells were clonally expanded and cryopreserved for drug tolerance studies. Cell lines were maintained at 37 °C in 5% CO₂. Media changes were performed every 2–3 days.
To generate early drug-tolerant cells, PC9 and/or SKMEL28 cells were plated on 100 mm plates (1 × 10⁵ cells/plate) and allowed to attach for 16 h. Media were replaced with fresh growth media containing gefitinib (Selleck Chemicals, Houston, TX, USA) (5 μM) or dabrafenib (Selleck Chemicals) (5 μM) for PC9 or SKMEL28 cells, respectively. Cells were cultured for the indicated amount of time, and media changes were performed every 2–3 days.

Celeste F.V, & Powers S. (2024). Induction of Multiple Alternative Mitogenic Signaling Pathways Accompanies the Emergence of Drug-Tolerant Cancer Cells. Cancers, 16(5), 1001.

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Cell Line Acquisition and Authentication

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Cell lines 293, HPAC, SW620, CT26, 4T1, and BALB/3T3 were obtained from the American Type Culture Collection (ATCC, cat nos. CRL-1573, CRL-2119, CRL-2638, CRL- 2539 and CCL-163, respectively). E0771 cells were obtained from CH3 BioSystems (cat. no. 940001) and PC9 cells were from Sigma (cat. no. 90071810). B16, glioma 261, and UACC-64 (UACC) cell lines were from the DCTD Tumor Repository at NCI (Frederick, MD). MC38, RENCA, and CHO-PR230 (CHO) cell lines were gifts of Jeffrey Schlom (NCI, NIH), Jonathan M. Weiss (NCI, NIH), and Stephen H. Leppla (National Institute of Allergy and Infectious Diseases [NIAID]), respectively. CHO-TEM8 and CHO-CMG2 cells were previously described^{21 (link)}, as were HCT-116-luc cells⁶⁵.

Hsu K.S., Dunleavey J.M., Szot C., Yang L., Hilton M.B., Morris K., Seaman S., Feng Y., Lutz E.M., Koogle R., Tomassoni-Ardori F., Saha S., Zhang X.M., Zudaire E., Bajgain P., Rose J., Zhu Z., Dimitrov D.S., Cuttitta F., Emenaker N.J., Tessarollo L, & St. Croix B. (2022). Cancer cell survival depends on collagen uptake into tumor-associated stroma. Nature Communications, 13, 7078.

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Cell Culture Protocols for Cancer and Normal Cell Lines

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NIH3T3 and HEK-293T cells were purchased from the American Type Culture Collection (ATCC) and grown as recommended in Dulbecco’s Modified Eagle Medium (DMEM) containing 10% FBS at 37°C in 5% CO₂. HCC2935, NCI-H460, NCI-H358, and NCI-H1437 cells were purchased from the American Type Culture Collection and grown as recommended in RPMI containing 10% FBS at 37°C in 5% CO₂, and PC9 cells were purchased From Sigma-Aldrich and grown as recommended in RPMI 10% FBS at 37°C in 5% CO_2. HSAEC1-KT cells were purchased from the ATCC and grown as recommended in SABM small airway epithelial cell-specific growth basal medium (Lonza Cat. No. CC-3119) along with singleQuots supplements and growth factors (Lonza Cat. No. CC-4124 (also see Key resources table).

Chava S., Bugide S., Zhang X., Gupta R, & Wajapeyee N. (2022). Betacellulin promotes tumor development and EGFR mutant lung cancer growth by stimulating the EGFR pathway and suppressing apoptosis. iScience, 25(5), 104211.

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Cell Culture Protocols for Various Cells

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The human embryonic kidney (HEK293T) and HeLa (CCL2) cells were obtained from the American Type Culture Collection and maintained in culture in DMEM supplemented with 10% FBS. H441 cells were obtained from the American Type Culture Collection and maintained in culture in RPMI medium with 10% FBS. PC9 cells were obtained from Sigma-Aldrich and maintained in culture in RPMI medium with 10% FBS. All cell lines were grown at 37 °C, 5% CO₂, and high humidity. All cell lines including stable clones were routinely evaluated for mycoplasma contamination using the LookOut Mycoplasma PCR Detection Kit (MP0035, Sigma-Aldrich) according to the manufacturer’s protocol.

Wisniewski D.J., Liyasova M.S., Korrapati S., Zhang X., Ratnayake S., Chen Q., Gilbert S.F., Catalano A., Voeller D., Meerzaman D., Guha U., Porat-Shliom N., Annunziata C.M, & Lipkowitz S. (2022). Flotillin-2 regulates epidermal growth factor receptor activation, degradation by Cbl-mediated ubiquitination, and cancer growth. The Journal of Biological Chemistry, 299(1), 102766.

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Culturing Diverse Cell Lines for EGFR Research

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LNCaP (CRL-1740), C4-2B (CRL-3315), T-47D (HTB-133), HEK293 (CRL-1573), and Phoenix-AMPHO (CRL-3213) cells, and SYF (CRL-2459) mouse embryonic fibroblasts were obtained from ATCC. PC-9 cells were obtained from Sigma. LNCaP and C4-2B cells were cultured in RPMI with 10% fetal bovine serum (FBS). PC-3 cells were cultured in F12K with 10% FBS. PC-9 cells were cultured in RPMI with 5% FBS. Human embryonic kidney 293 cells and SYF cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 10% FBS. EGF from mouse was purchased from Sigma (E5160). For EGFR inhibition, erlotinib (Thermo Fisher Scientific) is used at 1 μM for 1 h.

Alwanian W.M., Vlajic K., Bie W., Kajdacsy-Balla A, & Tyner A.L. (2022). Protein tyrosine kinase 6 regulates activation of SRC kinase. The Journal of Biological Chemistry, 298(11), 102584.

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Fluorescent Lung Cancer Cell Lines

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A-549 (CCL-185), NCI-H1299 (CRL-5803), NCI-H2228 (CRL-5935), NCI-H460 (HTB-177), NCI-H1734 (CRL-5891), NCI-H1993 (CRL-5909), and HCC827 (CRL-2868) cells were from ATCC. PC-9 cells were from Sigma-Aldrich. NCI-H3122 cells were from the National Cancer Institute. All cell lines were transduced with the FU-EBFP2-H2B-W plasmid that expresses a nuclear-localized blue-fluorescent protein. All cell lines were cultured in RPMI1640 supplemented with FBS (10% v/v), glutamine (4 mM), and penicillin/streptomycin (100 U/mL). PC9, H1229, and H3122 cell lines were also cultured in HPLM supplemented with FBS (10% v/v) and penicillin/streptomycin (100 U/mL).

Ghezzi C., Perez S., Ryan K., Wong A., Chen B.Y., Damoiseaux R, & Clark P.M. (2022). Early reduction of glucose consumption is a biomarker of kinase inhibitor efficacy which can be reversed with GLUT1 overexpression in lung cancer cells. Molecular imaging and biology, 25(3), 541-553.

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Cell culture conditions for cancer cell lines

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293T cells (CRL-3216), A549 cells (CCL-185), H1299 cells (CRL-5803), H1975 cells (CRL-5908) and SW620 cells (CCL-227) were purchased from ATCC. PC9 cells were purchased from Sigma Aldrich (9007180). A549 cells were cultured in complete F-12K medium (10% FBS, 2 mM glutamine, 100 μg ml⁻¹ penicillin and 100 μg ml⁻¹ streptomycin) at 37 °C in 5% CO₂. SW620 cells were cultured in complete Leibovitz’s L-15 Medium (10% FBS, 2 mM glutamine, 100 μg ml⁻¹ penicillin and 100 μg ml⁻¹ streptomycin) in 37 °C air with no CO₂. PC9, H1975 and H1299 cells were cultured in complete RPMI (RPMI 1640 supplemented with 2 mM l-glutamine, 1 mM sodium pyruvate, 1 mM nonessential amino acids, 20 mM HEPES pH 7.5, 50 mM b-mercaptoethanol, 100 U ml⁻¹ penicillin, 100 mg ml⁻¹ streptomycin, 10% FBS). 293T cells were cultured in complete DMEM (10% FBS, 2 mM glutamine, 100 μg ml⁻¹ penicillin and 100 μg ml⁻¹ streptomycin) at 37 °C in 5% CO₂.

Kubo S., Fritz J.M., Raquer-McKay H.M., Kataria R., Vujkovic-Cvijin I., Al-Shaibi A., Yao Y., Zheng L., Zou J., Waldman A.D., Jing X., Farley T.K., Park A.Y., Oler A.J., Charles A.K., Makhlouf M., AbouMoussa E.H., Hasnah R., Saraiva L.R., Ganesan S., Al-Subaiey A.A., Matthews H., Flano E., Lee H.H., Freeman A.F., Sefer A.P., Sayar E., Çakır E., Karakoc-Aydiner E., Baris S., Belkaid Y., Ozen A., Lo B, & Lenardo M.J. (2021). Congenital iRHOM2 deficiency causes ADAM17 dysfunction and environmentally directed immunodysregulatory disease. Nature immunology, 23(1), 75-85.

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Cell Culture Conditions for Cancer Research

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PC-9 cells were obtained originally from Sigma-Aldrich Corporation. HeLa cells, NIH3T3 cells, COS1 cells, H441 cells, H3225 cells, and H1975 cells were obtained originally from American Type Culture Collection. All cells were cultured in DMEM or RPMI 1640 medium containing 10% fetal bovine serum (FBS) within 5% CO₂ atmosphere at 37° C.

Yu C.H., Chou C.C., Tu H.F., Huang W.C., Ho Y.Y., Khoo K.H., Lee M.S, & Chang G.D. (2018). Antibody-assisted target identification reveals afatinib, an EGFR covalent inhibitor, down-regulating ribonucleotide reductase. Oncotarget, 9(30), 21512-21529.

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Establishment and Characterization of Isogenic EGFR-Resistant Cell Lines

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All cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. Sensitive and resistant cell lines were established from the same ancestral population of PC9 cells (Sigma-Aldrich, 90071810). Resistant population was cultured in 1 μM gefitinib (Cayman, 13166) for greater than 6 months until a population of stably growing cells was observed. Resulting subpopulations exhibited noticeable visual morphological differences in the culture. The sensitive population was cultured in parallel in matched volumes of DMSO (Sigma-Aldrich, 276855) for the same duration as a vehicle control.
Resulting resistant and sensitive subclones underwent lentiviral transduction with plasmid vectors encoding EGFP and mCherry fluorescent proteins with attached nuclear localization sequence (plasmids were a gift from A.M.’s laboratory at Moffitt Cancer Center). Derivative cell lines with heritable fluorescent protein expression were selected in puromycin (MP Biomedical, 100552).

Farrokhian N., Maltas J., Dinh M., Durmaz A., Ellsworth P., Hitomi M., McClure E., Marusyk A., Kaznatcheev A, & Scott J.G. (2022). Measuring competitive exclusion in non–small cell lung cancer. Science Advances, 8(26), eabm7212.

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