Jurkat e6 1

Manufactured by American Type Culture Collection

Sourced in United States

Jurkat E6-1 is a T-cell line derived from a human acute T-cell leukemia. It is a commonly used cell line in immunological research and is suitable for a variety of cell-based assays.

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18 protocols using jurkat e6 1

Characterization of Human T-ALL Cell Lines

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The human T-ALL cell lines Jurkat (E6.1), CCRF-CEM, HSB-2 and Molt-3 were from ATCC (Manassas, VA) and were cultured in RPMI 1640 medium containing 10% of fetal bovine serum, 100 units/ml penicillin and streptomycin, and 2 mmol/l glutamine (complete medium).
T-ALL patients were diagnosed and treated at Hôpital Saint-Louis (Paris, France). The patients or relatives have signed the consent forms according to the Declaration of Helsinki. Hôpital Saint-Louis and Institut Universitaire d’Hématologie Institutional Review Board approved the study. The study was performed with cryopreserved leukemic cells isolated from the blood of three stage IV T-ALL patients at diagnosis.

Berrazouane S., Boisvert M., Salti S., Mourad W., Al-Daccak R., Barabé F, & Aoudjit F. (2019). Beta1 integrin blockade overcomes doxorubicin resistance in human T-cell acute lymphoblastic leukemia. Cell Death & Disease, 10(5), 357.

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Human Cell Lines in RPMI 1640 Medium

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Human cell lines [RL, CEM, Jurkat E6.1, Jurkat A3, Jurkat I 9.2 (caspase-8 mutant)] were purchased from ATCC and maintained in RPMI 1640 medium (Life technologies) supplemented with 10% FBS (Life Technologies) at 37°C, 5% CO₂. RL cells B-lymphoblasts originally isolated from a patient with non-Hodgkin’s lymphoma, CEM cells are T-lymphoblasts originally isolated from a patient with acute lymphoblastic leukemia, and Jurkat cells are T-lymphocytes originally isolated from a patient with acute T-cell leukemia.

DiFranco K.M., Johnson-Farley N., Bertino J.R., Elson D., Vega B.A., Belinka BA J.r, & Kachlany S.C. (2015). LFA-1-targeting Leukotoxin (LtxA; Leukothera®) causes lymphoma tumor regression in a humanized mouse model and requires caspase-8 and Fas to kill malignant lymphocytes*. Leukemia research, 39(6), 649-656.

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Cell Line Maintenance and Genetic Modification

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The Jurkat E6-1 and K562 cell lines were obtained from ATCC and maintained in R10. The Capan-2, Panc-1, PL45, Hs766T, BT-20, MCF-7, MDA-MB-231, and MDA-MB-453 cell lines were also obtained from ATCC and maintained in DMEM supplemented with 10% FCS, penicillin and streptomycin. NNP4 primary ovarian cancer cells were obtained for a pleural tap of an ovarian cancer patient at the University of Chicago Hospitals. K562meso was generated through transduction of K562 cell line with lentiviral supernatant containing mesothelin cDNA expressed by the EF1α promoter. Luciferase-expressing cell lines were generated through transduction of the parental cell line with lentiviral supernatant containing Click Beetle Green luciferase-T2A-GFP and sorted for GFP expression on the BD Influx (BD Biosciences). Cell lines expressing COSMC were generated by transducing the parental cell lines with lentiviral supernatant containing CD19t-P2A-cosmc and sorted on the BD Influx for expression of CD19.

Posey AD J.r., Schwab R.D., Boesteanu A.C., Steentoft C., Mandel U., Engels B., Stone J.D., Madsen T.D., Schreiber K., Haines K.M., Cogdill A.P., Chen T.J., Song D., Scholler J., Kranz D.M., Feldman M.D., Young R., Keith B., Schreiber H., Clausen H., Johnson L.A, & June C.H. (2016). Engineered CAR T Cells Targeting the Cancer-Associated Tn-Glycoform of the Membrane Mucin MUC1 Control Adenocarcinoma. Immunity, 44(6), 1444-1454.

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HIV-1 Molecular Clone Cell Culture

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The HEK293T (ATCC) and TZM-bl (John C. Kappes, Xiaoyun Wu, and Tranzyme, Inc) cell lines were cultured in DMEM (Gibco) supplemented with 10% heat-inactivated FBS (Gibco), 2 mM glutamine, 100 U mL^–1 penicillin, and 100 μg mL^–1 streptomycin. The SupT1 (ATCC) and the Jurkat E6-1 and Jurkat CypA-/- (Douglas Braaten and Jeremy Luban) cell lines were cultured in RPMI 1640 medium supplemented with 10% heat-inactivated FBS, 2 mM glutamine, 100 U mL^–1 penicillin, and 100 μg mL^–1 streptomycin. Cells were cultured at 37°C with 5% CO₂. Viruses were generated from the full-length HIV-1 molecular clone pNL43 and its mutant derivatives. Details about the construction of the pNL43 molecular clones harboring CA mutations are included in the Supplementary Materials and Methods.

Balasubramaniam M., Davids B.O., Bryer A., Xu C., Thapa S., Shi J., Aiken C., Pandhare J., Perilla J.R, & Dash C. (2022). HIV-1 mutants that escape the cytotoxic T-lymphocytes are defective in viral DNA integration. PNAS Nexus, 1(2), pgac064.

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Cell Line Procurement and Validation

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HepG2 (ATCC HB-8065), Hep3B (ATCC HB-8064), HeLa (ATCC CLL-2), HL-60 (ATCC CLL-240), and Jurkat E6-1 (Aids reagent cat# 177) were purchased from the respective suppliers.

Setten R.L., Chomchan P., Epps E.W., Burnett J.C, & Rossi J.J. (2021). CRED9: a differentially expressed elncRNA regulates expression of transcription factor CEBPA. RNA, 27(8), 891-906.

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Cell Culture Protocols for Common Cell Lines

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Parental HEK293, Jurkat E6-1, RBL-1, DLD1, HCT116, and A20 cell lines were purchased directly from ATCC (Catalog # CRL-1573, TIB-152, CRL-1378, CCL-221, CCL-247, and TIB-208). HEK293, RBL-1, and HeLa cells were cultured in high-glucose (4.5 g/l) Dulbecco's modified Eagle's medium supplemented with 10% heat-inactivated fetal bovine serum and 1× Antibiotic–Antimycotic solution. Jurkat, A20, and DLD1 were maintained in RPMI 1640 with L-glutamine supplemented with 10% heat-inactivated fetal bovine serum and 1× Antibiotic–Antimycotic solution. HCT116 were cultured in McCoys 5A medium supplemented with 10% heat-inactivated fetal bovine serum and 1× Antibiotic–Antimycotic solution. All cell lines were stored in a heated (37 °C) humidified incubator under standard cell culture conditions (5% CO₂; 95% air). The absence of mycoplasmal contamination was verified for all cell lines using a sensitive PCR-based detection kit (ABM: G238).

Yoast R.E., Emrich S.M., Zhang X., Xin P., Arige V., Pathak T., Benson J.C., Johnson M.T., Abdelnaby A.E., Lakomski N., Hempel N., Han J.M., Dupont G., Yule D.I., Sneyd J, & Trebak M. (2021). The Mitochondrial Ca2+ uniporter is a central regulator of interorganellar Ca2+ transfer and NFAT activation. The Journal of Biological Chemistry, 297(4), 101174.

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Cell Culture Conditions for Multiple Cell Lines

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MDA-MB-231, MCF7, Jurkat E6.1 (European Collection of Authenticated Cell Cultures), HEK293, HL-60, and RAMOS (ATCC) were cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS), penicillin (50 units/mL), and streptomycin (50 µg/mL) in a humidified incubator at 37 °C with 5% CO₂. MCF10A breast epithelial cell line (ATCC) was cultured in mammary epithelial cell growth medium (MEGM) with MEGM SingleQuot Supplements (hydrocortisone, rhEGF, insulin, and bovine pituitary extract) as instructed (Lonza) and 100 ng/mL cholera toxin (Sigma). Cell lines were passaged by 0.05% trypsin and 0.5 mM EDTA (Gibco) and quenched with serum-containing medium. MCF10A cells were trypsinized and quenched with Defined trypsin Inhibitor (Thermo Fisher Scientific). Cell viability was measured by trypan blue exclusion assay. Briefly, 10 µL of cell suspension was mixed with 10 µL of 4% trypan blue solution and total and trypan-stained cells were quantified by Countess II Automated Cell Counter (Thermo Fisher Scientific).

Switzer C.H., Cho H.J., Eykyn T.R., Lavender P, & Eaton P. (2022). NOS2 and S-nitrosothiol signaling induces DNA hypomethylation and LINE-1 retrotransposon expression. Proceedings of the National Academy of Sciences of the United States of America, 119(21), e2200022119.

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Culture of T-ALL Cell Lines

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The human T-ALL cell lines Jurkat (E6.1) and HSB-2 were purchased from ATCC (Manassas, VA, USA) and were cultured in RPMI 1640 medium with 10% of fetal bovine serum (FBS), 100 units/mL penicillin, streptomycin and 2 mmol/L glutamine (complete medium).

Berrazouane S., Doucet A., Boisvert M., Barabé F, & Aoudjit F. (2021). VLA-4 Induces Chemoresistance of T Cell Acute Lymphoblastic Leukemia Cells via PYK2-Mediated Drug Efflux. Cancers, 13(14), 3512.

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Jurkat Cell Lines for PD-1 Signaling

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Jurkat E6-1 (ATCC), PD-1-NFAT luciferase
reporter Jurkat (BPS Bioscience) and PD-1-SNAP-Jurkat cells were maintained
in complete RPMI 1640 supplemented with 10% heat-inactivated fetal
bovine serum (ThermoFisher Scientific). The cell lines were routinely
screened for mycoplasma contamination (Eurofins Genomics).

Fang T., Alvelid J., Spratt J., Ambrosetti E., Testa I, & Teixeira A.I. (2021). Spatial Regulation of T-Cell Signaling by Programmed Death-Ligand 1 on Wireframe DNA Origami Flat Sheets. ACS Nano, 15(2), 3441-3452.

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Establishing Cell Line Cultures for Research

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KBM-7 WT cells were obtained from Haplogen. HEK293T and K562, Jurkat E6.1 and Raji cells were purchased from ATCC. LAMA-84 cells were purchased from DSMZ. BRD4 reporter clones (REDS) were generated by retroviral-based gene trap technology as previously described^{15 (link),16 (link)} using the human male near-haploid CML cell line KBM-7. REDS1, REDS15, REDS17 and KBM-7 WT cells were maintained in Iscove’s Modified Dulbecco’s Medium (IMDM, Gibco). Jurkat E6.1, Raji, K562 and LAMA-84 were maintained in RPMI 1640. HEK293T cells were maintained in Dulbecco’s Modified Eagles Medium (DMEM, Gibco). All media were supplemented with 10% Fetal Bovine Serum (FBS, Gibco) and 1% penicillin/streptomycin (PS). cell lines were maintained at 5% CO₂ at 37°C, initially authenticated by STR typing before use (except the REDS) and tested for mycoplasma contamination.

Li K.C., Girardi E., Kartnig F., Grosche S., Pemovska T., Bigenzahn J.W., Goldmann U., Sedlyarov V., Bensimon A., Schick S., Lin J.M., Gürtl B., Reil D., Klavins K., Kubicek S., Sdelci S, & Superti-Furga G. (2021). Cell-surface SLC nucleoside transporters and purine levels modulate BRD4-dependent chromatin states. Nature metabolism, 3(5), 651-664.

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