Cell culture media and hemoglobin were obtained from Sigma-Aldrich (St. Louis, MO, USA), fetal bovine serum (FBS) from Gibco (Rockville, MD, USA), bovine serum albumin (BSA) (Fraction V) from USB (Cleveland, OH, USA). J774A.1 cells were obtained from ATCC (catalog number TIB-67) and human coronary endothelial cells from Lonza (Basel, Switzerland). CD163 was obtained from R&D Systems, Inc. (Minneapolis, MN, USA). Common Laboratory Chemicals were purchased from Fisher Scientific or Sigma. Peptide was synthesized, per our amino acid sequence plan, by Chinese Peptides Company, (Hangzhou, China) and fully characterized by nuclear magnetic resonance (NMR), carbon, hydrogen and nitrogen (C,H,N) composition analyses and by MALDI/FAB mass spectrometry. 111InCl3 (Nordion Inc., Ottawa, ON, Canada) was supplied by Nuclear and Energy Research Institute (IPEN)/CNEN, (São Paulo, Brazil). HPLC analysis were performed in a Shimadzu VP series equipment (Shimadzu, Japan) connected to radiation detector Flow Scintillation Analyzer, Radiomatic 610RT (PerkinElmer, Boston, MA, USA).
J774a 1 cells
Manufactured by American Type Culture Collection
Sourced in United States
J774A.1 cells are a murine macrophage cell line derived from the BALB/c mouse. They are commonly used in research to study macrophage biology and function.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Hemoglobin, by Merck Group (1 mentions)
Cd163, by R&D Systems (1 mentions)
Cell culture media, by Merck Group (1 mentions)
N18 tg2 cells, by Leibniz Institute DSMZ (1 mentions)
Celltiter aqueous one solution cell proliferation assay, by Promega (1 mentions)
Nl20 cells, by American Type Culture Collection (1 mentions)
Tcs sp5 2 confocal laser scanning microscope, by Leica (1 mentions)
Wst 1, by Roche (1 mentions)
F 12k medium, by Corning (1 mentions)
Sh sy5y cell, by American Type Culture Collection (1 mentions)
Dulbecco modified eagle medium (dmem), by American Type Culture Collection (1 mentions)
Hek293t, by American Type Culture Collection (1 mentions)
Tib 67, by American Type Culture Collection (1 mentions)
Thiazolyl blue tetrazolium bromide, by Merck Group (1 mentions)
26 protocols using j774a 1 cells
1
Radiolabeled peptide characterization
2
Murine BMDM and J774A.1 Cell Culture
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Murine BMDMs and J774A.1 cells (ATCC, Manassas, VA, USA) were cultured in DMEM containing 10% FBS at 37°C in a humidified incubator supplemented with 5% CO2. In some experiments, BMDMs were cultured in glutamine-free DMEM medium for glutamine deprivation.
3
Cell Culture of J774A.1 and N18 TG2 Cells
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J774A.1 cells were purchased from ATCC (Manassas, VI, USA) and N18 TG2 cells from the DSMZ (Braunschweig, Germany), Pregnant OF1/SPF mice were purchased from the Institute for Laboratory Zoology and Veterinary Genetics, Himberg (Austria). All cells were cultured in an incubator at 37 °C, with 5% CO2, and 100% humidity.
4
Cytotoxicity Evaluation of Algae-NP-robot
5
Candida albicans Infection in Macrophages
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Mouse RAW264.7 cells and J774A.1 cells (ATCC, USA; Catalog#: TIB-71 and TIB-67) was cultured in high glucose Dulbecco’s Modified Eagle’s Medium (DMEM) (Invitrogen, USA; Catalog #: 220511) with 10% FBS (Gibco, USA; Catalog #: A31605), 100× penicillin-streptomycin (Thermo Fisher Scientific, USA; Catalog #: 10378016) at 37°C with 5% CO2. Before stimulating with C. albicans, the culture medium was replaced with fresh medium and 1 × 105 CFU of C. albicans suspended in DMEM was administered. The cells were cultured in the presence of C. albicans for 0, 1, 2, 6, 12, and 24 h. For co-treatment with C. albicans and miR-384-5p inhibitor, the RAW264.7 cells, and J774A.1 cells were treated with C. albicans for 24 h.
6
Characterizing Macrophage Lipid Uptake
7
Evaluating Anthrax Toxin Variants
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A mouse macrophage cell line (J774A.1 cells, ATCC TIB-67, Manassas, VA) were plated in 96-well flat-bottomed tissue culture plates at 2.5x104 cells/well in 50 μL and incubated for 16–19 hours at 37°C with 5% CO2. pp-PA83 variants were titrated on the plated cells in the presence or absence of LF (List Biological Laboratories, Campbell, CA). After a 4 hour incubation at 37°C with 5% CO2, cell viability was accessed by adding WST-1 (Roche Applied Sciences, Indianapolis, IN), a proliferation reagent, followed by a spectrophotometric measurement at 450 nm. A four-parameter logistic-log regression model was used to analyze the OD versus the PA83 concentration. The inflection point for each curve from this model is reported as showed the effective concentration 50% (EC50) in ng/mL of PA83 for the corresponding protein sample.
8
Maintenance of Cell Lines for Experiments
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HEK293 cells (American Type Culture Consortium) were maintained in complete media (Dulbecco’s Modified Eagle’s Medium (DMEM)) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37°C in 5% CO2. J774A.1 cells (female mouse macrophages, ATCC) were maintained in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin. SH-SY5Y cells, (ATCC) were maintained in a 1:1 mixture of F-12K Medium (Kaighn’s Modification of Ham’s F-12 Medium) (Corning Inc., New York, NY, USA) and DMEM supplemented with 10% FBS and 1% penicillin/streptomycin.
9
Culturing J774A.1 Macrophage Cells
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J774A.1 cells (ATCC, Manassas, VA, United States) were cultured in T25 flasks in Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, Thermo Fisher Scientific, Waltham, MA, United States) with 10% fetal bovine serum (Sigma-Aldrich, United States) and maintained at 37°C, 5% CO2. Cells were scraped using Teflon cell scraper from the T25 flasks and the final cell density of 2 to 4 × 105 cells/mL was adjusted using fresh DMEM. The cells were cultured for 48 h at 37°C until 60–80% confluent in a 6-well plate (each plate representing individual time point and concentration).
10
Culturing Mouse Macrophage J774.A1 Cells
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Mouse macrophage J774.A1 cells (American Type Culture Collection, Manassas, VA, USA) were maintained in Dulbecco's modified Eagle's medium containing 10% (v/v) fetal bovine serum, 100 U/mL penicillin, and 100 U/mL Fungizone under 100% humidity and 5% CO2, at 37°C.
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