E g7 ova

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E.G7-OVA is a cell line derived from the C57BL/6 mouse strain. It is an antigenic tumor cell line that expresses the ovalbumin (OVA) protein.

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Lab products found in correlation

Thp 1, by American Type Culture Collection (3 mentions) Hek293t, by American Type Culture Collection (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) L glutamine, by Thermo Fisher Scientific (3 mentions) Fetal bovine serum (fbs), by MP Biomedicals (3 mentions) Penicillin, by Thermo Fisher Scientific (3 mentions) Streptomycin, by Thermo Fisher Scientific (3 mentions) E g7 ova cell line, by UniProt (2 mentions) Mv4 11, by American Type Culture Collection (2 mentions) Pl 21, by American Type Culture Collection (2 mentions) Molm 13, by American Type Culture Collection (2 mentions) C1498, by American Type Culture Collection (2 mentions) Hepa1 6, by American Type Culture Collection (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Gentamycin, by Thermo Fisher Scientific (2 mentions) β mercaptoethanol, by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Mem nonessential amino acid, by Nacalai Tesque (2 mentions) Rpmi 1640, by Nacalai Tesque (2 mentions) L glutamine, by Fujifilm (2 mentions)

Spelling variants of this product

E.G7-OVA cells (19 citations) E.G7 OVA cell line (5 citations) EL4-OVA (E.G7-OVA), (2 citations) E.G7-OVA thymoma cells (2 citations) EL4 and E.G7-OVA cells (2 citations)

38 protocols using e g7 ova

Generating Cell Lines for Immunotherapy

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E.G7-OVA (derivative of EL4) cells were obtained from ATCC (Manassas, VA, Cat. # CRL-2113) and maintained via the ATCC-recommended culture methods. E.G7-OVA cells were lentivirally transduced to express PD-L1, as previously described.^{19 (link)} The A2/sarcoma cell line expressing SSX2 (A2/Sarc-SSX2) was generated as previously described.^{16 (link)} The MycCaP cell line was obtained from ATCC (Cat #CRL-3255) and cultured according to their instructions. All cell lines used were authenticated and tested for mycoplasma.

Zahm C.D., Moseman J.E., Delmastro L.E, & G. Mcneel D. (2021). PD-1 and LAG-3 blockade improve anti-tumor vaccine efficacy. Oncoimmunology, 10(1), 1912892.

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Culturing Immune-Modulating Cell Lines

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The TC-1 cell line (lung epithelial cells from C57BL/6 mice transformed with HPV16 E6/E7 and the c-Ha-ras oncogene) was kindly provided by Dr T-C Wu (John Hopkins University, USA). TC-1 cells were maintained in RPMI-1640 medium (HyClone) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (HyClone), 50 units/mL penicillin/streptomycin, 1 mM sodium pyruvate, 25 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, Biological Industries) and 55 µM β-mercaptoethanol. E.G7-OVA (ATCC, CRL-2113) is a derivative of mouse thymoma EL4 cells transfected with the ovalbumin (OVA) plasmid. The E.G7-OVA cells were cultured in RPMI-1640 medium supplemented with 10% (v/v) heat-inactivated fetal bovine serum, 50 units/mL penicillin/streptomycin, 1 mM sodium pyruvate, 20 mM HEPES, 50 µM β-mercaptoethanol and 0.4 mg/mL G418. B16F10-OVA melanoma cells were cultured in Dulbecco's Modified Eagle Medium (DMEM, Gibco) supplemented with 10% (v/v) heat-inactivated fetal bovine serum, 50 units/mL penicillin/streptomycin and 1 mg/mL G418. All cells were maintained at 37°C in 5% CO₂ and determined to be mycoplasma negative before the animal study.

Yan W.L., Wu C.C., Shen K.Y, & Liu S.J. (2021). Activation of GM-CSF and TLR2 signaling synergistically enhances antigen-specific antitumor immunity and modulates the tumor microenvironment. Journal for Immunotherapy of Cancer, 9(10), e002758.

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HTLV-1 Infected Cell Line Cultivation

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The HTLV-1-infected cell lines MT1, MT2, and MT4 were purchased from JCRB Cell Bank. The HTLV-1-infected cell line MJ, and the T-cell lymphoma cell lines Jurkat and EG7-OVA were provided by ATCC (VA, USA). All cell lines are identified based on short tandem repeat profiles by providers, and mycoplasma contaminations were denied both by providers and at our laboratories. Cell lines and PBMCs were cultivated using RPMI 1640 Medium (Nacalai Tesque, Kyoto, Japan) supplemented with 10% fetal bovine serum (FBS), and Penicillin–Streptomycin Mixed Solution (×100) (Nacalai Tesque). Mouse anti-CD4, CD8, CD7, CD39, CD73, and CD26 antibodies were purchased from Biolegend (San Diego, CA, USA), and anti-CADM1 antibody was purchased from MBL Inc. (Woburn, MA, USA). adenosine 5′-triphosphate disodium salt hydrate, adenosine 5′-monophosphate hydrate, and adenosine were purchased from Sigma Aldrich (now Merck KGaA, Darmstadt, Germany). Polyinosinic-polycytidylic acd sodium salt (Poly(I:C)) was purchased from R&D systems (Minneapolis, MN, USA). The CellTrace^TM Violet Cell Proliferation Kit was purchased from Thermo Fisher Scientific (Waltham, MA, USA).

Nagate Y., Ezoe S., Fujita J., Okuzaki D., Motooka D., Ishibashi T., Ichii M., Tanimura A., Kurashige M., Morii E., Fukushima T., Suehiro Y., Yokota T., Shibayama H., Oritani K, & Kanakura Y. (2020). Ectonucleotidase CD39 is highly expressed on ATLL cells and is responsible for their immunosuppressive function. Leukemia, 35(1), 107-118.

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Diverse Cell Lines for Cancer Research

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PL-21, THP-1, MOLM-13, MV4-11, E.G7-OVA, and SEM cell lines were purchased from ATCC (USA). The E.G7-OVA cell line was modified to express full-length human EGFRvIII (Uniprot Entry P00533 AA 1-29, 298–646), resulting in E.G7-EGFRvIII cells. Luciferase-eGFP (LUC-GFP) overexpressing cell lines PL-21-LUC-GFP, THP-1-LUC-GFP and MV4-11-LUC-GFP were generated according to previously described protocols [22 (link)]. Antigen quantification of cell lines are summarized in Supplementary Table 1A. 293Vec-Galv and 293Vec-RD114 were a kind gift of Manuel Caruso, Québec, Canada and have been previously described [27 (link)]. All human cell lines were short tandem repeat profiled in house to verify their origin. Cells were used for a time period no longer than two months.

Benmebarek M.R., Cadilha B.L., Herrmann M., Lesch S., Schmitt S., Stoiber S., Darwich A., Augsberger C., Brauchle B., Rohrbacher L., Oner A., Seifert M., Schwerdtfeger M., Gottschlich A., Rataj F., Fenn N.C., Klein C., Subklewe M., Endres S., Hopfner K.P, & Kobold S. (2021). A modular and controllable T cell therapy platform for acute myeloid leukemia. Leukemia, 35(8), 2243-2257.

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Murine EL4 Lymphoblast Cell Line for Tumor Studies

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The murine EL4 lymphoblast cell line expressing OVA (E.G7-OVA) was purchased from ATCC (ATCC Cat# CRL-223). Cells were cultured for two passages before being implanted in the left flank of naive female mice. Further authentication of the cell line was not performed. The sex of the cell line is female. Cells were maintained in 1640 RPMI media supplemented with 10% GIBCO serum, 1% Penstrep and 1% L-Glut and 55 μM beta-mercaptoethanol, under 5% CO₂ atmospheric oxygen, at 37°C in a humidified incubator.

Field C.S., Baixauli F., Kyle R.L., Puleston D.J., Cameron A.M., Sanin D.E., Hippen K.L., Loschi M., Thangavelu G., Corrado M., Edwards-Hicks J., Grzes K.M., Pearce E.J., Blazar B.R, & Pearce E.L. (2020). Mitochondrial Integrity Regulated by Lipid Metabolism Is a Cell-Intrinsic Checkpoint for Treg Suppressive Function. Cell Metabolism, 31(2), 422-437.e5.

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Evaluating Tumor Immunity in Mice

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EG.7-OVA, (ATCC) and was maintained in CM prior to subcutaneous injection into the left flank of B6 or TLR2^−/− mice. Tumors were allowed to progress to approximately 250 mm³ in volume prior to the addition of intravenous administration of 10⁶ indicated naïve CD4⁺ T cells. Tumors were evaluated for changes in tumor volume every three days with the formula 3.14 × [largest diameter × (perpendicular diameter) ²]/6. In some experiments mice were injected intraperitoneally with 1 mg/kg of a pan-specific neutralizing antibody against TGF-β1, -β2, and -β3 (R&D Systems) or control rabbit polyclonal IgG for one day before and every other day until the conclusion of the experiment. PD-L1 neutralization was conducted with intraperitoneal administration of 100 μg of the clone 10F.9G2 (Bio-X-Cell) or control mouse IgG every 3 days after tumor establishment until completion of the study. Tumors were minced with a razor, digested with DNAse I (Sigma) and Collagenase/Dispase (Roche) for one hour at 37 °C and cell suspensions were filtered through a 70 μm filter. Tumor cells and infiltrating lymphocytes were quantitated using fluorescent flow cytometric counting beads (BD Biosciences).

Ibrahim M., Scozzi D., Toth K.A., Ponti D., Kreisel D., Menna C., De Falco E., D’Andrilli A., Rendina E.A., Calogero A., Krupnick A.S, & Gelman A.E. (2017). Naïve CD4+ T cells carrying a TLR2 agonist overcome TGF–β-mediated tumor immune evasion. Journal of immunology (Baltimore, Md. : 1950), 200(2), 847-856.

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Characterization of Murine Tumor Models

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Unless otherwise noted, all reagents were purchased commercially and used as received. Oligonucleotides were synthesized as described below. Peptides were purchased from Genscript or Northwestern’s Peptide Synthesis core. Chemicals were purchased from suppliers listed in parentheses. C57BL/6 mice and C57BL/6-Tg(TcraTcrb)1100Mjb/J (OT-1, 003831) female mice, age 8–12 weeks old, were purchased from Jackson Laboratory. Mice were used in accordance with all national and local guidelines and regulations and protocols performed were approved by the institutional animal use committee at Northwestern University (IUCAC). E.G7-OVA and B16-F10 cells were purchased from ATCC. MC-38 cells were kindly provided by Dr. Bin Zhang. Antibodies were purchased and clones are provided in Supplementary Table 3.

Xie S., Green J., Bixby J.B., Szafranska B., DeMartini J.C., Hecht S, & Roberts R.M. (1997). The diversity and evolutionary relationships of the pregnancy-associated glycoproteins, an aspartic proteinase subfamily consisting of many trophoblast-expressed genes. Proceedings of the National Academy of Sciences of the United States of America, 94(24), 12809-12816.

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Establishing Genetically Modified Cell Lines

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PL-21, THP-1, MOLM-13, MV4-11, E.G7-OVA and SEM cell lines were purchased from ATCC (USA). The E.G7-OVA cell line was modified to express full-length human EGFRvIII (Uniprot Entry P00533 AA 1-29, 298-646), resulting in E.G7-EGFRvIII cells. Luciferase-eGFP (LUC-GFP) overexpressing cell lines PL-21-LUC-GFP, THP-1-LUC-GFP and MV4-11-LUC-GFP were generated according to previously described protocols (22 (link)). Antigen quantification of cell lines are summarized in Supplementary Table 1A. 293Vec-Galv and 293Vec-RD114 were a kind gift of Manuel Caruso, Québec, Canada and have been previously described (27 (link)). All human cell lines were short tandem repeat profiled in house to verify their origin. Cells were used for a time period no longer than two months.

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Adoptive T Cell Therapy for Tumor Models

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Female C57BL/6 mice aged 5–6 weeks were purchased from the Experimental Animal Center of Fujian Medical University (Fuzhou, China). Recipient CD45.1 mice and donor OT-I/CD45.2/Rag^−/− mice were bred in-house. The donor OT-I/CD45.2/Rag^−/− mice express a transgenic T cell receptor (TCR) that recognizes the H-2Kb-restricted class I epitope of ovalbumin (OVA_257−264, SIINFEKL). All mice were maintained in pathogen-free facilities at Fujian Medical University.
MC38-OVA and MC38 colorectal carcinoma cells were obtained from the laboratory of Dr. Lie-Ping Chen (Yale University). EG7-OVA and EL4 lymphoma cells were purchased from ATCC (Manassas, USA). All tumor cell lines were tested before use and found to be free of mycoplasma by PCR mycoplasma test kit (HuaAn Biotechnologies, China).

Lai J.Z., Zhu Y.Y., Ruan M., Chen L, & Zhang Q.Y. (2019). Local Irradiation Sensitized Tumors to Adoptive T Cell Therapy via Enhancing the Cross-Priming, Homing, and Cytotoxicity of Antigen-Specific CD8 T Cells. Frontiers in Immunology, 10, 2857.

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Generation of Bst2 Knockout and Transgenic Mice

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Tumor cell lines YAC-1 (ATCC TIB-160), RMA-S, B16, EL4 (ATCC TIB-39), and E.G7-OVA (ATCC CRL-2113) were cultured in Roswell Park Memorial Institute (RPMI) 1640 Medium (RPMI 1640, Gibco) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 1% penicillin-streptomycin, 10 μg/mL gentamycin, and 50 µM β-mercaptoethanol (Gibco-BRL). Bst2 knockout mice (C57BL/6 background) were created on a C57BL/6Tac background by Xenogen Biosciences (Cranbury, NJ). NZW/N mice expressing cytoplasmic tail deficient, short isoform of BST2 (Bst2^S/S) were purchased from Japan SLC (Hamamatsu, Japan) and crossed with Bst2 knockout C57BL/6 (B6) mice to generate B6/NZW F1 mice. To generate C57BL/6 mice containing Bst2 gene of NZW origin, NZW/N mice were backcrossed with C57BL/6 Bst2^-/- mice at least eight times. All animal experiment protocols of this study were approved by the Institutional Animal Care and Use Committee of Korea University (KUIACUC-2018-25, 29 March~31 December 2018; and KUIACUC-2019-0003, 1 January 2019~31 December 2020).

Oh J., Yi E., Jeong S.K., Park S, & Park S.H. (2022). BST2, a Novel Inhibitory Receptor, Is Involved in NK Cell Cytotoxicity through Its Cytoplasmic Tail Domain. International Journal of Molecular Sciences, 23(19), 11395.

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