The human erythroleukemia cell lines YN-1 [27 (link)] and K562 [28 (link)], human myeloid leukemia cell line KG1a [29 (link)], human T cell leukemia cell line Jurkat, human monocytoid cell line U937, and the human pre-B cell leukemia cell line NALM6 were cultured in RPMI-1640 (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS; Biowest, Miami, FL, USA) and 1% penicillin/streptomycin (Sigma-Aldrich). Stable YN-1 cells expressing pGL4.20 (GATA-2 +9.9/1S; described below) were cultured in RPMI-1640 containing 10% FBS, 1% penicillin/streptomycin, and 1 μg/ml puromycin (Sigma-Aldrich). K562 cells were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA); other cell lines (YN-1, KG1a, Jurkat, U937, and NALM6) were obtained from the Cell Resource Center for Biomedical Research at Tohoku University (www2.idac.tohoku.ac.jp/dep/ccr/ /).
Jurkat
Manufactured by Merck Group
Sourced in United States
Jurkat is a human T lymphocyte cell line derived from the peripheral blood of a patient with T cell leukemia. It is commonly used in immunological and cell signaling research as a model system for studying T cell function and activation.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640, by Merck Group (4 mentions)
Penicillin streptomycin, by Merck Group (3 mentions)
Rpmi 1640 medium, by Merck Group (3 mentions)
Mcf 7, by Merck Group (2 mentions)
Ethylene glycol tetraacetic acid (egta), by Merck Group (2 mentions)
Jurkat clone e6 1, by American Type Culture Collection (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Jurkat, by American Type Culture Collection (2 mentions)
K562 cell, by American Type Culture Collection (1 mentions)
Puromycin, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Dmem medium, by Merck Group (1 mentions)
Mda mb 231, by Merck Group (1 mentions)
Sk br 3, by Merck Group (1 mentions)
Hepes, by Merck Group (1 mentions)
Flt3l, by Thermo Fisher Scientific (1 mentions)
Recombinant human il 3, by Thermo Fisher Scientific (1 mentions)
Stemspan sfem 2 medium, by STEMCELL (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
10 protocols using jurkat
1
Culturing Human Leukemia Cell Lines
2
Cultivation of Human Cancer Cell Lines
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Human breast adenocarcinoma cell lines (MCF7 [RRID: CVCL_0031], SKBR3 [RRID:CVCL_0033], BT549 [RRID:CVCL_1092], and MDA-MB231 [RRID:CVCL_0062]), human non-small cell lung cancer cell line (A549) [RRID:CVCL_0023], human colorectal carcinoma cell line (HCT116) [RRID:CVCL_0291], human histiocytic lymphoma cell line (U937) [RRID:CVCL_0007], human acute T cell leukemia cell line (Jurkat) [RRID:CVCL_0367], Abelson murine leukemia cell line (Raw 246.7) [RRID:CVCL_0493] and human normal fibroblast cell line (F-180) were obtained from the Radiobiology and Experimental Radio Oncology lab, University Cancer Center, Hamburg University, Hamburg, Germany. Doxorubicin-resistant MCF7 and A549 cell lines were generated in our lab. MCF7, SKBR3, MDA-MB231, and A549 cells were maintained in RPMI-1640 medium (Sigma-Aldrich-St. Louis, MO, USA). Media were supplied with 10% fetal bovine serum (Sigma-Aldrich-St. Louis, MO, USA) and 1% penicillin/streptomycin (Sigma-Aldrich-Louis, MO, USA). HCT116, U937, Jurkat, Raw 246.7, and F-180 were cultured in DMEM medium (Sigma-Aldrich-St. Louis, MO, USA), which is supplied with 10% fetal bovine serum (Sigma-Aldrich-St. Louis, MO, USA) and 1% penicillin/streptomycin (Sigma-Aldrich-St. Louis, MO, USA). All incubations were performed at 37 °C in a humidified atmosphere of 5% CO2.
3
Jurkat Cells Culture and Electrophysiology
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The human leukemic T cell line, Jurkat (clone E6-1), was purchased from American Type Culture Collection (Manassas, VA). The Jurkat cells were grown in RPMI 1640 medium (Sigma-Aldrich, St. Louis, MO) containing 10 % heat-inactivated FBS, 10 mM HEPES and 2 mM glutamate. Cells were grown on culture plates at 37 °C in a 5 % CO2-humidified incubator. During the experiments, cells were placed in the external solution containing, in mM: 150 NaCl, 4.5 KCl, 1 CaCl2, 1 MgCl2, 10 HEPES and pH 7.35 adjusted with NaOH, 300 mOsm. The pipette solution contained in mM: 150 KCl, 1 CaCl2, 2 MgCl2, 10 HEPES 10 EGTA, and pH 7.2 was adjusted with KOH, 280 mOsm. The concentration of free calcium ions in the internal solution was below 100 nM, assuming the dissociation constant for EGTA at pH 7.2 of 10−7 M (Grissmer et al. 1993 (link)). Such a low-calcium concentration was applied to prevent the activation of calcium-activated K+ channels KCa2.2 abundantly expressed in Jurkat T cells (Grissmer et al. 1992 (link)). The chemicals were purchased from the Polish Chemical Company (POCH, Gliwice, Poland), except of HEPES and EGTA that were purchased from SIGMA. The examined compounds were purchased from Alexis Biochemicals (Lausen, Switzerland).
4
Culturing Hematopoietic Cell Lines and Primary JMML Cells
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Hematopoietic cell lines PEER, K-562, MONO-MAC-6, HL-60, OCI-AML3, JURKAT, DAUDI, KASUMI-1, LOUCY, MV-4-11, and THP-1 were available in house, DEL and JVM-2 were purchased at the DSMZ repository (Braunschweig, Germany), and KARPAS-422 at Sigma-Aldrich (Saint Louis, Missouri, USA). Cell lines were grown in RPMI medium (Invitrogen, Waltham, MA, USA) supplemented with 10% or 20% Fetal Calf Serum (FCS, ThermoFisher Scientific, Waltham, MA, USA), according to supplier instructions, together with 100 U/mL Penicillin/Streptomycin (10,000 U/mL, Invitrogen) and 100 µg/mL l -glutamine (200 mM, Invitrogen). For THP-1, medium was additionally supplied with 0.05 mM β-mercaptoethanol. Cell lines were incubated at 37 °C in 5% CO2 incubators.
Mononuclear cell preparations derived from spleen and PB from 3 JMML patients were primary cultured in StemSpan SFEM II medium (Stemcell Technologies, Vancouver, Canada) supplemented with recombinant human IL3 (0.01 µg/mL; PeproTech, London, UK), FLT3L (0.01 µg/mL; PeproTech, London, UK), TPO (0.01 µg/mL; PeproTech, London, UK) and SCF (0.025 µg/mL; PeproTech, London, UK) and incubated at 37 °C in 5% CO2 incubators.
Mononuclear cell preparations derived from spleen and PB from 3 JMML patients were primary cultured in StemSpan SFEM II medium (Stemcell Technologies, Vancouver, Canada) supplemented with recombinant human IL3 (0.01 µg/mL; PeproTech, London, UK), FLT3L (0.01 µg/mL; PeproTech, London, UK), TPO (0.01 µg/mL; PeproTech, London, UK) and SCF (0.025 µg/mL; PeproTech, London, UK) and incubated at 37 °C in 5% CO2 incubators.
5
In Vitro Culture of Human Cancer Cell Lines
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Five human cancer cell lines—liver (HepG2), breast (MCF-7), prostate (PC-3 and LNCap), and blood (Jurkat), obtained from RIKEN BioResource Centre Cell Bank (Japan) were used for this study. The cell lines were cultured as previously described,16 (link) in RPMI 1640 (PC-3, LNCap, and Jurkat) and Dulbecco’s modified Eagle medium (DMEM) (MCF-7, HepG2) medium supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich, Urbana, IL, USA) and 1% of penicillin streptomycin L-glutamine (Sigma-Aldrich, Urbana, IL, USA). Cells were grown in a humidified incubator (Panasonic Healthcare Company Limited, Japan) at 37 °C, supplied with 5% CO2 and passaged on reaching about 90% confluency.
6
Cell Line Cultivation and Maintenance
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Human embryonic kidney (HEK) 293T, HeLa, and the human hematopoietic cell lines K562 and Jurkat, were obtained from the American Type Culture Collection (ATCC). EBV transformed B-lymphoblast cells (LCL) were generated by immortalizing primary B lymphocytes with EBV by the Diagnostic Immunology Laboratory, CCHMC. Leukocyte adhesion deficiency (LAD) patient-derived B lymphocytes cells were EBV immortalized and were obtained from Dr. Dennis Hickstein at the National Cancer Institute (NIH). HeLa EJ5-GFP cells with an integrated I-SceI GFP reporter cassette for NHEJ assessment were kindly provided by Dr. Younghoon Kee at the University of South Florida. HeLa cells, K562 cells, and 293T cells were cultured in DMEM (Corning) supplemented with 10% fetal bovine serum (FBS; VWR), 1% l -glutamine (MP Biomedicals) and 1% Penicillin-Streptomycin (Lonza). LAD patient-derived B lymphocytes, Jurkat, and LCL cells were cultured in RPMI-1640 (Sigma) supplemented with 20% FBS and 1% Penicillin-Streptomycin.
7
Jurkat T Cell Electrophysiology Protocol
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The human leukemic T cell line, Jurkat (clone E6–1), was purchased from American Type Culture Collection (Manassas, VA). The Jurkat cells were grown in RPMI 1640 medium (Sigma-Aldrich, St. Louis, MO) containing 10% heat-inactivated FBS, 10 mM HEPES and 2 mM glutamate. Cells were grown on culture plates at 37 °C in a 5% CO2-humidified incubator. During the experiments cells were placed in the external solution containing in mM: 150 NaCl, 4.5 KCl, 1 CaCl2, 1 MgCl2, 10 HEPES: pH of solution =7.35. The pipette solution contained in mM: 150 KCl, 1 CaCl2, 2 MgCl2, 10 HEPES, 10 EGTA, pH of solution =7.2. The concentration of free calcium ions in the internal solution was below 100 nM, assuming the dissociation constant for EGTA at pH=7.2 of 10−7 M [21] (link). Such a low calcium concentration was applied to prevent the activation of calcium-activated K+ channels KCa2.2 abundantly expressed in Jurkat T cells [22] (link). The chemicals were purchased from the Polish Chemical Company (POCH, Gliwice, Poland), except of HEPES and EGTA that were purchased from SIGMA. The examined statins were purchased from Alexis Biochemicals (Lausen, Switzerland).
8
Human Cancer Cell Line Culture
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The human cancer cell lines Jurkat (acute T-cell leukemia, ATCC, Manassas, VA), A2780 (ovarian carcinoma, Sigma Aldrich, St. Louis, MO, US), HeLa (cervical carcinoma, ATCC), U2OS (osteosarcoma, ATCC), Hep3B (hepatoma, ATCC), and LN229 (glioblastoma, ATCC) were used during this study. Jurkat and A2780 cells were grown in RPMI-1640 (Sigma Aldrich), HeLa, Hep3B, and LN229 cells were cultured in Dulbecco’s modified eagle medium (DMEM, Sigma Aldrich). All growth media were supplemented with 10% heat-inactivated fetal bovine serum (FCS, PAA, Biowest, Nuaillé, France). Cells were passaged twice a week and regularly tested on Mycoplasma contamination (Mycoplasma kit, Sigma Aldrich). Cells were kept in 37 °C humid incubators containing 5% CO2 and 10% O2. For indicated experiments, the O2 level was reduced to 0–0.5%, creating a hypoxic environment.
9
Cell Culture Maintenance Protocol
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Jurkat (ATCC Cat# TIB-152, RRID : CVCL_0367), HeLa (ATCC Cat# CCL-2.2, RRID : CVCL_0058), NOMO-1 (DSMZ Cat# ACC-542, RRID : CVCL_1609), K562 (ATCC Cat# CCL-243, RRID : CVCL_0004) and U87 (ATCC Cat# HTB-14, RRID : CVCL_0022) cell lines were all acquired from ATCC and cultured as recommended by the supplier. LAN-1 cell line (DSMZ Cat# ACC-655, RRID : CVCL_1827) was acquired from DSMZ and cultured as recommended by the supplier. Cell lines were screened monthly for mycoplasma contamination Briefly, Jurkat, K562 and NOMO-1 cells were grown in 10% FBS-supplemented RPMI1640 (Sigma Aldrich) suspension culture and kept at <1x106/mL density. LAN-1, HeLa and U87 cell lines were grown in 10% FBS-supplemented DMEM (Thermo Fisher) adherent culture and split regularly at around 80-90% confluency using trypsin (Thermo Fisher)-based disaggregation, to avoid overgrowth.
10
Cell Line Culture Protocol
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Five established cell lines were included in the study: 3 CTCL cell lines (Hut‐78—Sézary syndrome, ATCC TIB‐161; HH—aggressive cutaneous T‐cell leukaemia/lymphoma, ATCC CRL‐2105; and SeAx—Sézary syndrome, kindly provided by Dr Markus Möbs13 ), T‐cell acute lymphoblastic leukaemia (T‐ALL) cell line Jurkat (SIGMA 88042803), and T‐cell Hodgkin lymphoma cell line HDLM2 (DSMZ ACC17). They were cultured in a HEPES‐buffered RPMI1640 medium with L‐glutamine (Thermo Fisher Scientific™), 10%‐20% foetal bovine serum (Sigma) and 1% penicillin/streptomycin (Life Technologies), according to manufacturer's instructions. Medium for SeAx was supplemented with Il‐2 (200 U/mL) (Sigma‐Aldrich) and medium for Jurkat with 1% sodium pyruvate (1 mmol\L) and 0.25% glucose (0.5 g/L) (Life Technologies).
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