HTLV-1-infected T-cell lines (ED-40515(−), MT-2, Hut-102, and ATL-43T) and non–HTLV-1–infected T-cell line (Jurkat T cell) were cultured in RPMI1640 (Nacalai Tesque) containing 10% FBS and 1% PSG (Invitrogen). Jurkat T cells stably expressing a spliced form of HBZ called Jurkat-HBZ were maintained as described previously44 (link). HEK293T cells were maintained with DMEM (Nacalai Tesque) containing 10% FBS and 1% PSG. The Jurkat Tet/On-Tax cells45 (link) kindly gifted from Dr. W.C. Greene were maintained in RPMI1640 supplemented with 10% tetracycline-free FBS (Clontech) and treated with 1 µg/ml doxycycline for 36 hours for inducible Tax expression. Human-cell-killing assay was performed as described previously9 (link).
Rpmi 1640
Manufactured by Takara Bio
Sourced in Japan
RPMI 1640 is a cell culture medium that supports the growth and maintenance of a variety of cell types, including mammalian cells, lymphocytes, and hybridomas. It is a widely used and well-established medium that provides a balanced combination of amino acids, vitamins, salts, and other essential components to support cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Tetracycline free fbs, by Takara Bio (1 mentions)
Penicillin streptomycin glutamine (psg), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin glutamine (psg), by Nacalai Tesque (1 mentions)
Rpmi 1640, by Nacalai Tesque (1 mentions)
Dulbecco modified eagle medium (dmem), by Nacalai Tesque (1 mentions)
Complete dmem, by Merck Group (1 mentions)
Lenti x 293t, by Takara Bio (1 mentions)
Hek293t, by American Type Culture Collection (1 mentions)
Heat inactivated fcs, by Merck Group (1 mentions)
Penicillin streptomycin, by Corning (1 mentions)
Rpmi 1640, by Corning (1 mentions)
Penicillin streptomycin, by Takara Bio (1 mentions)
Sybr green pcr master mix, by Takara Bio (1 mentions)
Tapi 0, by Enzo Life Sciences (1 mentions)
Total rna isolation kit, by Takara Bio (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Fetal calf serum (fcs), by Cytiva (1 mentions)
Charcoal stripped fbs, by Merck Group (1 mentions)
Tet system approved fbs, by Takara Bio (1 mentions)
8 protocols using rpmi 1640
1
HTLV-1-Infected T-Cell Lines and Assays
2
Maintenance of Diverse Cell Lines
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Cell lines were maintained at 37°C with 5% CO2 in a humidified incubator (Heraeus). Human embryonic kidney epithelial 293T cells HEK293T (ATCC CRL-3216) and Lenti-X293T (Clontech) cell lines were cultured in complete DMEM (Sigma Aldrich) supplemented with 15% fetal bovine serum (FBS). Jurkat-derived cell line variants 31.13 (Alcover et al., 1990 (link)) (abbreviated J31.13) and J76 (Heemskerk et al., 2003 (link)), which lack expression of TCRβ and TCRαβ respectively, were cultured in RPMI 1640 supplemented with 10% FBS. Key resources table lists all the above cell lines and those that were derived from J31.13 and J76 in this study. Cells were routinely tested and found negative for mycoplasma and are not STR profiled. Jurkat cell lines containing a tetracycline-inducible gene expression systems were maintained in RPMI 1640 supplemented with 10% tetracycline-negative FBS (Clontech).
3
Ovarian Cancer Cell Line Characterization and Culture
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OVCA429 parental cell line and OVISE cell line expressing tetracycline-inducible wild-type ARID1A (17 (link)) originated from the laboratory of I. M. Shih. Cell lines were tested for mycoplasma (University of Pennsylvania School of Medicine, Cell Culture Services). All cell lines were cultured on polystyrene in a 2D format in the presence of 5% CO2 at 37 °C. OVCA429 cells were maintained in RPMI 1640 (Corning, Corning, NY, cat. no. 10–092-CM) supplemented with 10% heat-inactivated FCS (Sigma-Aldrich, St. Louis, MO, cat. no. F4135). OVISE cells expressing inducible wild-type ARID1A were maintained in RPMI 1640 supplemented with 10% Tet System Approved FCS (Clontech, Mountain View, CA, cat. no. 631107). All media contained 1% Penicillin-Streptomycin (Corning, cat. no. 30-002-CI).
4
Isolation and Characterization of Astragaloside Compounds
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Chemically pure astragaloside total (AST) and astragaloside IV (ASIV) were purchased from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China) and their purity is greater than 98%. Stock solutions of AST and ASIV were made in DMSO. The final concentration of DMSO added to cells was never greater than 0.1%. Recombinant mouse TNFα was obtained from Calbiochem (San Diego, CA, USA). Cycloheximide (CHX) was purchased from Sigma (St. Louis, MO, USA). TAPI-0 (TNF-α processing inhibitor-0), a synthetic inhibitor of TNFα converting enzyme (TACE), was purchased from Enzo Life Sciences Inc (Farmingdale, NY, USA). RPMI 1640, glutamine, penicillin/streptomycin, and HEPES were purchased from Clontech Laboratories (Palo Alto, CA, USA). Fetal calf serum (FCS) was purchased from Hyclone Laboratories (Logen, UT, USA). Total RNA isolation kits, reverse transcription (RT) kits and SYBR Green PCR Master MIX were all purchased from Takara Bio Inc. (Dalian, China). Cell Surface Protein Isolation Kits were purchased from Pierce (Rockford, IL, USA).
5
Cultivation of KSHV-infected Cell Lines
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293T cells (kind gift of Lori Frappier, University of Toronto) were grown at 37°C in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and glutamine. iSLK cells [58 (link)] (gift of Don Ganem, UCSF) were maintained in DMEM containing 10% charcoal stripped FBS (Sigma) and 1% glutamine with 250μg/ml G-418 and 1μg/ml puromycin. iSLK cells were infected with WT KSHV derived from bacmid BAC16, expressing eGFP and hygromycin resistance [28 (link)]. Bac16 KSHV infected iSLK cells were maintained in 1.2 mg/ml hygromycin, 250μg/ml G-418 and 1μg/ml puromycin. TRExBCBL1-Rta (kindly provided by Prof. J. Jung) were cultured in RPMI 1640 10% Tet System approved FBS (Clontech) and 1% glutamine with 50μg/ml hygromycin.
6
Cultivating Lung Adenocarcinoma Cell Lines
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The human lung adenocarcinoma cell line A549 and the DDP-resistant cell line A549/DDP were purchased from the Tumor Cell Bank of the Chinese Academy of Medical Science and maintained in RPMI 1640 (TAKARA, Japan) containing 10% heat-inactivated fetal bovine serum (GIBCO, USA) in a humidified incubator with 5% CO2 at 37°C. A549/DDP cells were cultured in the presence of 10μM DDP (Yunnan Biovalley Dengzhanhua Pharmaceutical Co. Ltd, China), which was withdrawn for two generations before the experiments were performed. These cell lines grew in monolayers and were passaged when the cultures were 70-80% confluent.
7
Immune Cell Phenotyping and Proliferation
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Cetuximab was purchased from Merck Millipore (Darmstadt, Germany). The antibodies used in the present study were mouse anti-human CD3-fluorescein isothiocyanate (FITC; monoclonal; 1:100; cat. no. 55539; BD Biosciences, Franklin Lakes, NJ, USA), mouse anti-human CD56-phycoerythrin (PE) (monoclonal; 1:100; cat. no. 555516; BD Biosciences); human immunoglobulin G (hIgG; polyclonal; 1:200; cat. no. bs-0297P; Beijing Biosynthesis Biotechnology Co., Ltd., Beijing, China), rabbit anti-human EGFR (polyclonal; 1:200; cat. no. Sc-03AC; Santa Cruz Biotechnology, Inc.) rabbit anti-human Ki-67 (monoclonal; 1:200; cat. no ZA-0502; OriGene Technologies, Inc., Beijing, China). The PV-9000 polymer detection system for immunohistological staining, apoptosis detection kits in situ and diaminobenzidine color reagent were purchased from OriGene Technologies, Inc. The WST-1 cell proliferation reagent was purchased from Roche Diagnostics (Basel, Switzerland). RPMI-1640 and 0.25% ethylenediaminetetraacetic acid pancreatin were purchased from Takara Bio, Inc. (Otsu, Japan), and Ficoll-paque lymphocyte separation medium was purchased from GE Healthcare Life Sciences (Chalfont, UK). Recombinant IL-2 was purchased from Beijing Four Rings Biopharmaceutical Co., Ltd. (Beijing, China). Cells were analyzed by Moflow XDP flow cytometry with Summit version 5.2 software (Beckman Coulter, Inc., Brea, CA, USA).
8
Culturing Breast Cancer SK-BR-3 Cells
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A breast cancer cell line, SK-BR-3 cells, was cultured in RPMI 1640 (Takara Biomedical Technology, Japan) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, USA), penicillin (100 IU/ml; Sigma, USA), and streptomycin (100 µg/ml; Sigma, USA).
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