The T24 cell line, which originated from advanced high-grade bladder cancer, was purchased from ATCC (Manassas, VA, USA) and maintained in RPMI-1640 growth medium (Nissui, Tokyo, Japan) supplemented with 10% FBS (ICN Biomedicals, Aurora, OH, USA), 100 U/mL penicillin and 100 μg/mL streptomycin (Gibco, Grand Island, NY, USA) in a standard humidified incubator at 37°C in an atmosphere of 5% CO2. A stock solution of ALA (Sigma-Aldrich, St. Louis, MO, USA) was prepared in deionized water at a concentration of 10 mM and stored at −20°C. PPIX (Wako Pure Chemical Industries, Osaka, Japan) was dissolved in dimethylsulfoxide and used as a control solution.
Fetal bovine serum (fbs)
Manufactured by ICN Biomedicals
Sourced in United States, Japan
Fetal bovine serum (FBS) is a complex mixture of proteins, growth factors, and other components derived from the blood of bovine fetuses. It is commonly used as a supplement in cell culture media to support the growth and proliferation of various cell types.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by Thermo Fisher Scientific (6 mentions)
Penicillin, by Thermo Fisher Scientific (5 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (4 mentions)
Trypsin edta solution, by Merck Group (3 mentions)
Rpmi 1640 growth medium, by Nissui Pharmaceutical (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by ICN Biomedicals (2 mentions)
Penicillin, by ICN Biomedicals (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Nacalai Tesque (2 mentions)
Dulbecco s modified eagle s medium dmem, by Merck Group (2 mentions)
Dichloromethane, by Fujifilm (2 mentions)
L cystine dihydrochloride, by Merck Group (2 mentions)
Amino acid free dmem, by Nacalai Tesque (2 mentions)
100 w v trichloroacetic acid solution tca, by Fujifilm (2 mentions)
1 mol l hydrochloric acid hcl, by Fujifilm (2 mentions)
Reduced glutathione gsh, by Fujifilm (2 mentions)
Hepes buffered saline solution, by Lonza (2 mentions)
Penicillin streptomycin glutamine (psg), by Thermo Fisher Scientific (2 mentions)
Trypsin 0.04 edta, by Merck Group (1 mentions)
28 protocols using fetal bovine serum (fbs)
1
Photosensitizer-based Bladder Cancer Assay
2
Isolation and Culture of Human Gingival and Periodontal Ligament Fibroblasts
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Gingival tissue and periodontal ligament tissue was collected, finely chopped, and cultured, and the outgrowth cells of each were used as hGFs and hPLFs, respectively. Alpha-minimum essential media (α-MEM) (SIGMA) containing 10% fetal bovine serum (FBS) (ICN Biomedicals Inc., Solon, OH, USA) and 2% penicillin–streptomycin (Pn-St) (SIGMA, St. Louis, MO, USA) were used as serum and cultured at 37 °C in the presence of 5% CO2. A subculture was performed using 0.1% trypsin–0.04% EDTA (SIGMA).
3
A549 Cell Culture Protocol
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A549 human lung cancer cells were obtained from American Type Culture Collection. The A549 cells were maintained in RPMI 1640 Medium (Life technologies, Carlsbad, CA, USA) containing 10% fetal bovine serum (FBS; ICN Biomedicals, Aurora, OH, USA), 1 mM L-glutamine (Life Technologies), 100 units/ml penicillin, and 100 μg/ml streptomycin in a humidified atmosphere of 95% air and 5% CO2 at 37°C.
4
Endoglin DNA Vaccine Delivery via Attenuated Salmonella
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The attenuated Salmonella Typhimurium SL7207 (aroA¯) strain was provided by Dr. C. A. Guzmán (German Research Center of Biotechnology, Braunschweig, Germany). Bacteria were cultured in LB broth supplemented with 100 μg/mL of ampicillin and with a mixture of amino acids. Endoglin-based DNA vaccine (Salmonella Typhimurium SL7207 strain carrying pcDNA3.1(+) plasmid with inserted endoglin (ENG) coding sequence) was used [42 (link)]. Plasmid pBCMGSNeo carrying a gene encoding murine IL-12 was obtained from Dr. H. Yamamoto (Osaka University, Osaka, Japan). Plasmid preparations were isolated using QIAGEN-Endo Free Giga Kit (QIAGEN GmbH, Hilden, Germany). B16-F10 cells (murine melanoma, ATCC, Manassas, VA, USA) were maintained using RPMI 1640 medium (Gibco BRL, Paisley, UK) supplemented with 10% FBS (ICN Biomedicals, Costa Mesa, CA, USA). Cell cultures were maintained under standard conditions (37°C, 5% CO2, 95% humidity) (see Jarosz et al. [42 (link)]).
5
Isolation of Primary Mixed Glial Cells
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The primary mixed glial cells were prepared from neonatal C57BL/6J mouse pups as reported previously (Ma et al., 2011 (link)). In brief, after carefully removing the meninges, the neonatal brain was disintegrated by pipetting. The cell suspension was seeded in 10-cm culture dishes at a density of one brain per dish. Dulbecco's modified eagle medium (DMEM, 10 mL; Sigma, St. Louis, MO, USA) containing 10% fetal bovine serum (FBS; ICN Biomedicals, Aurora, OH, USA) was added to each dish. After14 days in vitro, primary mixed glial cells were dissociated by trypsinization, and the cell suspension was plated in 15.6-mm culture dishes for treatment with Cat C (R&D Systems, Minneapolis, MN, USA); each culture dish contained 1 × 106 cells.
6
Cell Line Maintenance Protocol
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The HCT116, HT29, LoVo, WiDr, CoLo201, and LS180 cell lines were obtained from the American Type Culture Collection (Manassas, VA). All cells were maintained in RPMI1640 medium supplemented with 10% fetal bovine serum (FBS) (ICN Biomedicals, Irvine, CA), 100 U/mL of penicillin G, and 100 μg/mL of streptomycin (Invitrogen Corp., Carlsbad, CA) in a humidified atmosphere of 5% CO2 at 37°C.
7
Toxicity Evaluation of H2O2 and RNSP in SH-SY5Y Cells
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Cells of the SH-SY5Y human neuroblastoma cell line, which were purchased from American Type Culture Collection (Manassas, VA, USA), were cultured in DMEM/F-12 mixture supplemented with 10% fetal bovine serum (FBS, ICN Biomedicals, Eschwege, Germany), 2 mM L-glutamine, and 1% antibiotic and antimycotic solution (Sigma, St. Louis, MO, USA) in a humid atmosphere of 5% CO2 and 95% air at 37°C. To determine the toxicity of the reagents, the cells were treated with freshly prepared H2O2 (from 30% stock) at concentrations ranging from 5 to 500 μM (diluted with sterile purified water), and RNSP was tested at concentrations ranging from 0.01 to 100 ug/mL (diluted with methanol or sterile purified water) for different treatment times.
8
Cultivation of Bladder Cancer Cell Lines
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Two human bladder cancer cell lines, T24 (ATCC no. HTB-4 derived from an undifferentiated grade 3 carcinoma) and MGH-U3 (a generous gift from Dr. H. LaRue at Laval University Cancer Research Centre, Quebec, Canada; derived from a grade 1 tumor), were maintained in RPMI-1640 growth medium (Nissui Pharmaceutical Co., Ltd.) supplemented with 10% fetal bovine serum (FBS) (ICN Biomedicals, Inc.), 100 U/ml penicillin, and 100 µg/ml streptomycin (Gibco; Thermo Fisher Scientific, Inc.) in a standard humidified incubator at 37°C in a 5% CO2 atmosphere for 24 h.
9
Modulation of Gefitinib Sensitivity in NSCLC Cells
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PC-9 and RPC-9 cells were kind gifts from Dr Kiura (Okayama University, Japan). A549, PC-9 and RPC-9 cells were cultured in RPMI 1640 Medium (Invitrogen, Carlsbad, CA, USA) with 10% FBS (ICN Biomedicals, Aurora, OH, USA), 2 mM l -glutamine (Life Technologies, Gaithersburg, MD, USA), 100 units/mL penicillin and 100 μg/mL streptomycin in 5% CO2 at 37°C. For preparation of PC-9 HGF cells, PC-9 cells were treated with 100 ng/mL HGF for 24 h. PC-9, RPC-9 or PC-9 HGF cells were treated with 10–10,000 nM gefitinib, 20 μM LY294002, 10 μM U0126, 10 μM SB203580 or 50/100 μM NSC23766 for the indicated time.
For knockdown experiments, 50 nM ON-target plus SMART pool siRAC1 or negative control #2 (Thermo Scientific, Rockford, IL, USA) were transfected in PC-9 cells using Lipofectamine RNAiMAX reagent (Life Technologies) and the transfected cells were subjected to a migration assay or western blotting after 72 h.
For transient transfection, pcDNA3.1-HA/RAC1G12V, -HA/RAC1Q61L or vector control plasmids with pEGFP-C1 (Clontech, Palo Alto, CA, USA) were co-transfected into PC-9 cells using Lipofectamine 2000 reagent (Life Technologies) and the transient transfected cells were subjected to a migration assay, western blotting or phalloidin staining after 48 h.
For knockdown experiments, 50 nM ON-target plus SMART pool siRAC1 or negative control #2 (Thermo Scientific, Rockford, IL, USA) were transfected in PC-9 cells using Lipofectamine RNAiMAX reagent (Life Technologies) and the transfected cells were subjected to a migration assay or western blotting after 72 h.
For transient transfection, pcDNA3.1-HA/RAC1G12V, -HA/RAC1Q61L or vector control plasmids with pEGFP-C1 (Clontech, Palo Alto, CA, USA) were co-transfected into PC-9 cells using Lipofectamine 2000 reagent (Life Technologies) and the transient transfected cells were subjected to a migration assay, western blotting or phalloidin staining after 48 h.
10
Culturing Metastatic RCC ACHN Cells
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ACHN cells were originally derived from malignant pleural effusion of a patient with widely metastatic RCC and were purchased from Dainipponseiyaku Co. (Tokyo, Japan). The ACHN cells were maintained in DMEM containing glucose (4.5 g/L) (Gibco-Invitrogen, Rockville, MD, USA) supplemented with 10% heat-inactivated FBS (ICN Biomedicals, Aurora, OH, USA) in 5% CO 2 incubators at 37°C.
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