The NHL cell lines JEKO-1, GRANTA-519, REC-1, MAVER-1, CA-46, DG-75, and SU-DHL-10 were purchased from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany), while MINO was obtained from ATCC (American Type Culture Collection, Manassas, VA, USA). The colon carcinoma cell line HCT116 wild type (HCT116 WT) was obtained from Dr. Bert Vogelstein (Johns Hopkins Cancer Center, Baltimore, MD, USA) and the HCT116 Bax/Bak double knockout (HCT116 Bax/Bak DKO) were kindly provided by Dr. Richard J. Youle (National Institutes of Health, Bethesda, MD, USA). All lymphoma cell lines were cultured in RPMI supplemented with 10% (GRANTA-519, REC-1, MINO) or 20% FCS (JEKO-1, MAVER-1) and 100 U/ml penicillin plus 100 µg/ml streptomycin. The HCT116 WT and HCT116 Bax/Bak DKO were cultured in DMEM supplemented with 10% FCS and 100 U/ml penicillin plus 100 µg/ml streptomycin.
Dg 75
Manufactured by Leibniz Institute DSMZ
Sourced in Germany
The DG-75 is a laboratory centrifuge designed for a wide range of applications. It is capable of processing various sample types at high speeds. The core function of the DG-75 is to separate and concentrate materials of different densities through the application of centrifugal force.
Automatically generated - may contain errors
Lab products found in correlation
Ca 46, by Leibniz Institute DSMZ (2 mentions)
Granta 519, by Leibniz Institute DSMZ (1 mentions)
Rec 1, by Leibniz Institute DSMZ (1 mentions)
Jeko 1, by Leibniz Institute DSMZ (1 mentions)
Su dhl 10, by Leibniz Institute DSMZ (1 mentions)
Maver 1, by Leibniz Institute DSMZ (1 mentions)
Powerplex 16 hs system, by Promega (1 mentions)
Q vd oph, by Selleck Chemicals (1 mentions)
1640 medium, by Thermo Fisher Scientific (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Iscove s modified dulbecco s medium, by Thermo Fisher Scientific (1 mentions)
Lenti x 293t cells, by Takara Bio (1 mentions)
Su dhl 4, by Leibniz Institute DSMZ (1 mentions)
Cho k1, by Leibniz Institute DSMZ (1 mentions)
Penicillin, by Lonza (1 mentions)
Fetal calf serum (fcs), by Harvard Bioscience (1 mentions)
Penicillin and streptomycin, by Harvard Bioscience (1 mentions)
Rpmi 1640 media, by Harvard Bioscience (1 mentions)
Gsk 872, by Merck Group (1 mentions)
6 protocols using dg 75
1
Cell Lines Sourcing and Culturing
2
Profiling B-cell lymphoma cell lines
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BL cell lines were purchased from ATCC (ST486) and DSMZ (CA46 and DG75); P493-6 B cells were a kind gift of Prof. D. Eick (Helmholtz center Munich). Cell lines were cultured at 37 • C under an atmosphere containing 5% CO2 in RPMI-1640 medium supplemented with 2 mM ultra-glutamine, 100 U/mL penicillin, 0.1 mg/mL streptomycin and 20% (ST486) or 10% (P493-6, CA46 and DG75) fetal calf serum (all reagents from Cambrex Biosciences, Walkersville, MD, USA). At regular intervals, cell lines were tested for mycoplasma contamination by PCR (61) and subjected to short tandem repeat genotyping using the PowerPlex 16HS system (Promega, Madison, WI, USA) to confirm cell line identity. The cell lines included in the panel were purchased from DSMZ, cultured according to their proposed culture conditions and included: PMBL-Primary mediastinal B-cell lymphoma (K1106P, MedB1); HL-Hodgkin lymphoma (L428, L540, L1236, KM-H2, SUP-HD1, DEV), DLbcL (SUDHL-2, -4 to -6 and -10, SC-1, OCILy3, U2932, DOHH2) and BL (ST486, CA46, DG75, RAMOS, BL65, Namalwa, Raij, Jijoye).
Q-VD-Oph (Cat# S7311; Selleckchem, Munich, Germany) stock of 5 mM dissolved in 100% DMSO was used to inhibit caspase activity. Medium was supplemented with Q-VD (10uM) or DMSO starting 24 h after lentiviral infection and on every following culture day.
Q-VD-Oph (Cat# S7311; Selleckchem, Munich, Germany) stock of 5 mM dissolved in 100% DMSO was used to inhibit caspase activity. Medium was supplemented with Q-VD (10uM) or DMSO starting 24 h after lentiviral infection and on every following culture day.
3
Culturing Cell Lines for Research
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Carnaval, DG-75 and SU-DHL-4 (German Collection of Microorganisms and Cell Cultures GmbH, DSMZ, Braunschweig, Germany) cells were cultured in Roswell Park Memorial Institute 1640 medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal calf serum (FCS, Thermo Fisher Scientific) and 1% penicillin (Pen)/streptomycin (Strep) solution (Lonza, Basel, Switzerland). Granta 519 (DSMZ), Chinese hamster ovary (CHO)-K1 (DSMZ) and Lenti-X™ 293T cells (Clontech, Saint-Germain-en-Laye, France) were kept in Dulbecco’s Modified Eagle’s Medium (Thermo Fisher Scientific) containing 10% FCS and 1% Pen/Strep. MEC2 cells (DSMZ) were maintained in Iscove’s Modified Dulbecco’s Medium (Thermo Fisher Scientific) supplemented with 20% FCS and 1% Pen/Strep. Cells were cultured in a humidified atmosphere at 37°C and 6% CO2.
4
Evaluating combinatorial effects of BL and DLBCL cell lines
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Human Burkitt’s lymphoma (BL) cell lines DG-75 and RAJI and diffuse large B-cell lymphoma (DLBCL) cell lines SU-DHL-4 (GCB) and U-2946 (ABC) were purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany), cultured as recommended by the manufacturer in RPMI 1640 media (Biochrom, Berlin, Germany) supplemented with 10% heat-inactivated FCS (Biochrom, Berlin, Germany) and 100 µg/mL penicillin and streptomycin (Biochrom, Berlin, Germany). Cultured cells were regularly tested for mycoplasma contamination and authenticity (cell surface markers by flow cytometry).
AZD5153 (AZD), I-BET 151 (GSK1210151A) (I-BET) and Entospletinib (GS-9973) (Ento) were obtained from Selleck Chemicals (Absource Diagnostics GmbH, Munich, Germany) and prepared according to the manufacturer’s instructions to a 10 mM stock and stored at −80 °C until ready for use.
Cell lines (3.33 × 10 5 cells/mL) were exposed to AZD, I-BET, Ento or DMSO (control) as mono substance (0.001 µM–10 µM) or in combination for up to 72 h (0.01 µM AZD, 0.1 µM I-BET, 1 µM Ento)
AZD5153 (AZD), I-BET 151 (GSK1210151A) (I-BET) and Entospletinib (GS-9973) (Ento) were obtained from Selleck Chemicals (Absource Diagnostics GmbH, Munich, Germany) and prepared according to the manufacturer’s instructions to a 10 mM stock and stored at −80 °C until ready for use.
Cell lines (3.33 × 10 5 cells/mL) were exposed to AZD, I-BET, Ento or DMSO (control) as mono substance (0.001 µM–10 µM) or in combination for up to 72 h (0.01 µM AZD, 0.1 µM I-BET, 1 µM Ento)
5
Generating Burkitt Lymphoma Cell Line for Infection Studies
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The human Burkitt lymphoma line DG75 was purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany). The cells were stably transfected with the cDNA of murine cationic amino-acid transporter 1 (slc7a1) to make them susceptible for infection with MMLV-based retrovirus particles and were selected with blasticidin S (10 μg ml−1). The resultant DG75EB cells were used in all experiments. DG75 cells deficient for BTK or GRB2, respectively, are described in detail in Supplementary Figs and were transfected wtih slc7a1 alike. Ramos cells expressing murine γ2am variants were previously described6 (link). All cells were cultured in RPMI1640+Glutamaxx (Biochrome) supplemented with 10% heat-inactivated FCS and antibiotics.
6
Characterization of Burkitt Lymphoma Cell Lines
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Human BL cell lines RAMOS, DG-75, RAJI, BJAB and DAUDI were obtained from the German Collection of Microorganisms and Cell cultures (DSMZ, Braunschweig, Germany) and BL-2, BL-30, Seraphine, BL-60, BL-70 and Salina cells were kindly provided by T. Oellerich, Department of Medicine II, Hematology/Oncology, University Hospital Frankfurt, Germany. All cell lines were authenticated by STR profiling and continuously monitored for mycoplasma contamination. Cells were cultured in RPMI 1640 (Life Technologies), supplemented with 10% or 20% fetal calf serum (FCS) and 1% penicillin/streptomycin (Invitrogen). The bivalent Smac mimetic BV6 was kindly provided by Genentech. The broad-range caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp(O-Me) fluoromethylketone (zVAD.fmk) was purchased from Bachem, Necrosulfonamide (NSA) from Toronto Research Chemicals Inc., GSK’872 and Necrostatin-1s (Nec-1s) from Merck and Dabrafenib from Selleck Chemicals. Recombinant human TRAIL was obtained from R&D Systems, human recombinant TNFα from PeproTech and human multimeric FASL from AdipoGen. Doxycycline hydrochloride (DOX) was purchased from Sigma-Aldrich. LCL-161 was purchased from Novartis and AT-406, Birinapant and SGI-110 (Guadecitabine) were obtained from Selleck Chemicals. All other chemicals were purchased from Sigma-Aldrich or Carl Roth unless indicated otherwise.
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