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RAW 264.7 Cells

RAW 264.7 Cells: A murine macrophage cell line derived from the ascites of a tumor induced by the Abelson murine leukemia virus.
These cells are commonly used in cell biology research to study macrophage function, inflammation, and immune response.
RAW 264.7 cells are a valuable model system for investigating cellular processes and testing therapeutic interventions relted to macrophage biology.

Most cited protocols related to «RAW 264.7 Cells»

1
Murine Norovirus Plaque Assay
RAW 264.7 cells were seeded into six-well plates at a density of 2 × 106 viable cells per well. On the following day, 10-fold dilutions of virus inoculum were prepared in complete DMEM medium and plated in duplicate wells. Plates were incubated for 1 h at room temperature on a rocking apparatus before aspirating the inoculum and overlaying the cells with 2 ml of 37–40 °C 1.5% SeaPlaque agarose in MEM supplemented with 10% low-endotoxin fet al.calf serum, 1% HEPES, 1% penicillin/streptomycin, and 2% glutamine (complete MEM) per well. Plates were incubated at 37 °C and 5% CO2 for 2 d. To visualize plaques, cells were stained with 2 ml of 56 °C 1.5% SeaKem agarose in complete MEM containing 1% neutral red per well for 6–8 h.
For plaque neutralization assays, differing concentrations of purified mAb (A6.2, anti-MNV-1 capsid; isotype control, 10H2, anti-reovirus μ1c) were incubated with equal plaque-forming units of either MNV-1.CW1 or MNV-1 brain homogenate for 30 min at 37 °C prior to performing the MNV-1 plaque assay.
Wobus C.E., Karst S.M., Thackray L.B., Chang K.O., Sosnovtsev S.V., Belliot G., Krug A., Mackenzie J.M., Green K.Y, & Virgin HW I.V. (2004). Replication of Norovirus in Cell Culture Reveals a Tropism for Dendritic Cells and Macrophages. PLoS Biology, 2(12), e432.
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Publication 2004
Biological Assay Brain Capsid Proteins Cells Endotoxins Glutamine HEPES Immunoglobulin Isotypes Orthoreoviruses Penicillins RAW 264.7 Cells Senile Plaques Sepharose Serum Streptomycin Technique, Dilution Virus
2
MTT Assay for Cell Viability
The cells were plated in 24 well culture plates (25000 cells/500 µl/well for all the cell lines except Raw264.7, which was seeded with 40,000 cells/well density) and incubated in CO2 incubator. Next day, treatment was given according to the experimental requirement. Further, at respective time points, 50 µl MTT solutions from the Stock (5 mg/ml) was added and cells were incubated in CO2 incubator in the dark for 2 hrs. The medium was removed and Formazan crystals formed by the cells were dissolved using 500 µl of DMSO followed by transfer in 96 well plate. The absorbance was read at 570 nm using 630 nm as reference wavelength on a Multiwell plate reader (Biotech Instruments, USA). Reduced formazan quantification was done with Formazan standard. At each respective time points cell numbers were counted with a Neubauer-improved counting chamber (Paul Marienfeld GmbH & Co. KG, Germany) under 10X objective, and 10X eye piece magnification with compound light microscope (Olympus CH30, Japan). After MTT incubation Formazan images were captured under bright field (20X objective) with inverted microscope (Olympus, Japan). All the parameters were recorded under similar experimental conditions.
Rai Y., Pathak R., Kumari N., Sah D.K., Pandey S., Kalra N., Soni R., Dwarakanath B.S, & Bhatt A.N. (2018). Mitochondrial biogenesis and metabolic hyperactivation limits the application of MTT assay in the estimation of radiation induced growth inhibition. Scientific Reports, 8, 1531.
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Publication 2018
Cells Formazans Light Light Microscopy Microscopy MTT formazan RAW 264.7 Cells Sulfoxide, Dimethyl
3
Evaluating Anti-Inflammatory Potential of RGSF
NO and cell viability assays were performed as described during our previous work [18] (link). Briefly, RAW264.7 cells (1×106 cells/mL) were pre-incubated with RGSF (25, 50, 100, and 200 μg/mL) or vehicle for 30 min and then stimulated with LPS (100 ng/mL) for 18 h. One-hundred microliter of cell supernatant from each well were transferred into 96-well microplates and mixed with an equal volume of Griess reagent at room temperature. The absorbance at 540 nm was determined by a Spectramax 250 microplate reader. For cell viability test, 30 μL of 5 mg/mL 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) reagent was added to the culture plates and cell viability test was performed based on the reduction of MTT reagent into an insoluble, dark purple formazan product in viable cells.
Yayeh T., Jung K.H., Jeong H.Y., Park J.H., Song Y.B., Kwak Y.S., Kang H.S., Cho J.Y., Oh J.W., Kim S.K, & Rhee M.H. (2012). Korean Red Ginseng Saponin Fraction Downregulates Proinflammatory Mediators in LPS Stimulated RAW264.7 Cells and Protects Mice against Endotoxic Shock. Journal of Ginseng Research, 36(3), 263-269.
Publication 2012
Biological Assay Bromides Cells Cell Survival Formazans Griess reagent RAW 264.7 Cells
4
Characterization of RAW 264.7 Macrophage Cell Line
The RAW 264.7 cell lines (third passage) were purchased from American Type Culture Collection (ATCC® TIB-71™) and from Sigma-Aldrich (cat. no. 91062702). Cells were cultured in DMEM high glucose medium supplemented with 10% FBS (all the experiments were performed using the same FBS batch) and 1% penicillin/streptomycin, in atmosphere of 5% CO2 and 95% humidity at 37°C. Cells were passaged after reaching 90% confluence, detached with cell scraper and subcultivated in 1:6 ratio in T-75 flasks. All cell culture equipment (flasks, pipettes etc.) used in this study were from the same batch. Cells were cultured continuously from the third passage until passage no. 50. Cells were frozen every fifth passage starting from the passage number three. Before phenotype and functional analysis cells were thawed and cultured for next two passages (e.g. cells of passage number three were thawed and cultured until passage no. 5 and then subjected to further analyses). Cells were regularly tested for Mycoplasma contamination. To avoid differences between culture techniques, cells were cultured only by one person.
Taciak B., Białasek M., Braniewska A., Sas Z., Sawicka P., Kiraga Ł., Rygiel T, & Król M. (2018). Evaluation of phenotypic and functional stability of RAW 264.7 cell line through serial passages. PLoS ONE, 13(6), e0198943.
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Publication 2018
Atmosphere Cell Culture Techniques Cells Cultural Evolution Culture Media Freezing Gastrin-Secreting Cells Glucose Humidity Mycoplasma Penicillins Phenotype RAW 264.7 Cells Streptomycin
5
Evaluation of Anti-inflammatory Effects

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Publication 2016
Cyclooxygenase-2 DNA, Complementary Glyceraldehyde-3-Phosphate Dehydrogenases Murine Leukemia Virus NOS2A protein, human Oligonucleotide Primers RAW 264.7 Cells Reverse Transcriptase Polymerase Chain Reaction RNA-Directed DNA Polymerase Tumor Necrosis Factor-alpha

Most recents protocols related to «RAW 264.7 Cells»

1
Mint Essential Oils Anti-Inflammatory Effects
Not available on PMC !

Example 7

The potential anti-inflammatory activity of mint essential oils (EOs) was assessed by nitric oxide (NO) production on LPS-stimulated murine RAW264.7 macrophages. The cell viability of macrophages with EO treatment was examined in parallel. LM-EO and KWM-EO show inhibitory activity on NO production in the LPS-stimulated RAW264.7 cells with IC50 values of 86.5 and 95.8 μg/mL, respectively (FIG. 8). Meanwhile, both mint essential oils have little or no detectable cytotoxicity to RAW264.7 cells at the same measured concentrations up to 100 μg/mL.

US11872260B2. Mint essential oils inhibit HRASQ61L mutant keratinocyte activity and prevent skin carcinogenesis (2024-01-16). ACADEMIA SINICA [TW]. Inventors: Lie-Fen Shyur [TW], Chih-Ting Chang [TW], Yuhsin Chen [TW].
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Patent 2024
Action Potentials Anti-Inflammatory Agents Cell Survival Cytotoxin Macrophage Mentha Mus Oils, Volatile Psychological Inhibition RAW 264.7 Cells
2
Antiviral Properties of Probiotic Compounds
Not available on PMC !

Example 10

CFSM was initially applied to virus-free cell cultures to determine the concentration of CFSM that could be used on FRhK cells without itself causing any detrimental effects on the cells. In FIGS. 17A and 17B, it can be seen that 2% CFSM is a desirable amount to use and this amount was used for subsequent experiments.

Next, viral particles in infected RAW 264.7 cells and media in the presence of probiotic CFSM were quantified. Mouse macrophage RAW 264.7 cells were infected with MNV-1 at 1×106 cells with 3.5×106 PFU. After RNA extraction from cells and media (supernatant), quantification of MNV-1 particles was done by a 2-step real-time PCR. The analysis showed a statistical difference (t-test, p<0.05) between the amounts of viral particles present in the media (FIG. 18, panel B) compared to untreated infected cells. For the number of viral particles inside the cells, only Lactococcus lactis and Lactobacillus reuteri (FIG. 18, panel A), showed a statistical difference compared to infected cells, however, Lactobacillus acidophilus La-5 showed a trend towards statistical significance. These results show that the propagation of MNV-1 might be negatively affected by the presence of bioactive compounds produced by probiotic strains.

US11857581B2. Antiviral methods and compositions comprising probiotic bacterial molecules (2024-01-02). MicroSintesis Inc. [CA]. Inventors: Mansel Griffiths [CA].
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Patent 2024
Cells Lactobacillus acidophilus Lactobacillus reuteri Lactococcus lactis Macrophage Mus Norovirus Infection Probiotics RAW 264.7 Cells Real-Time Polymerase Chain Reaction Strains Virion Virus Vision
3
Anti-inflammatory Effects of Apple Stem Cell Extracts
Not available on PMC !

Example 6

In an inflammatory reaction, activated cells (such as macrophages) release a variety of pro-inflammatory cytokines (such as tumor necrosis factor alpha (TNF-α). The released cytokines can be assayed as a measure of inflammatory activity. To evaluate the anti-inflammatory role of apple stem cell extracts, mouse RAW 264.7 cell lines mouse macrophages were used as an adherent monolayer on petri dishes. These cells could be harvested easily without damage caused by enzymes or cell scrapers. The macrophages were stimulated in suspension with lipopolysaccharide (LPS) to initiate an inflammatory response. Cells were seeded into 12-well cell culture plates containing the apple stem cell extract treatment materials. After 16-18 hours, the medium conditioned by the macrophages was harvested and the cytokine profile in the medium determined with enzyme-linked immunosorbent assays (ELISA) by measuring TNF-α levels.

Method: Three concentration of ASC (6.25, 12.5 and 25 μg/mL in media) were tested for the anti-inflammatory effect. RAW 264.7 mouse macrophage cells were maintained in DMEM containing Glutamax supplemented with 10% FBS, penicillin (100 U/ml) and streptomycin (100 μg/ml). The macrophages treated with LPS (1:500 dilution of a 0.1 mg/ml solution of LPS in phosphate buffered saline (PBS)) to produce a pro-inflammatory response. The ASC treatment was performed with a final concentration of 1×105 macrophages in wells of a 12-well plate. The cytokine assay was performed using a TNF-α ELISA from R&D Systems of Minneapolis, Minnesota.

Results indicated (Table 6, FIG. 5) that LPS alone produced an inflammatory response more than 1000 times that of unstimulated cells as measured by TNF-α expression. Treatment with ASC on the induced macrophages showed a dose-dependent decrease of TNF-α expression. ASC concentrations of 6.25, 12.5, and 25 μg/mL reduced TNF-α activity in the induced cells by 72.1, 92.1 and 94.5%, respectively. This reduced TNF-α at doses of 12.5 and 25 μg/ml was statistically significant with p≤0.05 for 25 μg/ml and p≤0.02 in 12.5 μg/ml. The apple stem cell extracts thus exerted an anti-inflammatory effect on the activated macrophage cells.

TABLE 6
Results of TNF-α release assay showing anti-inflammatory effects of apple
stem cell extracts on mouse RAW 264.7 macrophage cell line cells.
Values shown are averages of three sets of experiments.
ASC extracts dramatically reduced inflammatory responses in the target
cells, as exemplified by reduced TNF-α release
(greater inhibition of inflammation).
Apple Stem percent
Cell Extract TNF-α inhibition
Conc. (μg/ml)(pg/ml)vs. LPS
25481.8994.5
12.5687.992.1
6.252432.8972.1
LPS8712.630
unstimulated6.45

US11872258B2. Oral cavity treatments (2024-01-16). None [US]. Inventors: Maryam Rahimi [US].
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Patent 2024
Anti-Inflammatory Agents Biological Assay Cell Culture Techniques Cell Extracts Cells Cytokine Enzyme-Linked Immunosorbent Assay Enzymes Hyperostosis, Diffuse Idiopathic Skeletal Inflammation Lipopolysaccharides Macrophage Mus Penicillins Phosphates Psychological Inhibition RAW 264.7 Cells Saline Solution Stem, Plant Stem Cells Streptomycin Technique, Dilution Tumor Necrosis Factor-alpha
4
Cellular Uptake and Cytotoxicity of Cy5.5-Labeled Nanoparticles
The cellular uptake of Cy5.5-CNPs was investigated in three types of normal cells, such as breast cancer cells (4T1), rat cardiomyocytes (H9C2), mouse fibroblasts (L929) and macrophages (Raw264.7). Each cell was incubated with Cy5.5-CNPs (100, 225 or 900 μg/ml) for 4 or 24 h. After treatment, cells were fixed with 4% paraformaldehyde for 15 min, and stained with 4′,6-diamidino-2-phenylindole (DAPI) for 10 min. The cellular uptake was observed using a Leica TCS SP8 confocal laser-scanning microscope (CLSM; Leica Microsystems GmbH; Wetzlar, Germany). Quantitative analyses of the fluorescence images were performed using ImageJ software (NIH, Bethesda, MD, USA). The cytotoxicity was assessed by the Cell Counting Kit-8 (CCK-8) assay. Briefly, 5 × 104 4T1, H9C2, L929 and Raw264.7 cells were seeded into 96-well cell culture plates. Then, each cell was treated with different concentration of Cy5.5-CNPs ranging from 0 to 900 μg/ml. After 24 h of incubation, the cells were further incubated with cell culture medium containing 10% of CCK-8 solution for 20 min. Finally, the cell viability was analyzed using a microplate reader (VERSAmaxTM; Molecular Devices Corp., USA) with a wavelength of 450 nm.
Chang H., Yhee J.Y., Jeon S., Shim M.K., Yoon H.Y., Lee S, & Kim K. (2023). In vivo toxicity evaluation of tumor targeted glycol chitosan nanoparticles in healthy mice: repeated high-dose of glycol chitosan nanoparticles potentially induce cardiotoxicity. Journal of Nanobiotechnology, 21, 82.
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Publication 2023
Biological Assay Breast Carcinoma Cell Culture Techniques Cells Cell Survival Culture Media CY5.5 cyanine dye Cytotoxin Fibroblasts Fluorescence Macrophage Medical Devices Microscopy, Confocal Mus Myocytes, Cardiac paraform RAW 264.7 Cells
5
Osteoclast Differentiation in RAW264.7 Cells

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Publication 2023
Cell Nucleus Cells Macrophage Mus Osteoclasts RAW 264.7 Cells TRANCE Protein

Top products related to «RAW 264.7 Cells»

1
Fetal bovine serum (fbs) by Thermo Fisher Scientific
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
2
Dulbecco modified eagle medium (dmem) by Thermo Fisher Scientific
Sourced in United States, China, United Kingdom, Germany, France, Australia, Canada, Japan, Italy, Switzerland, Belgium, Austria, Spain, Israel, New Zealand, Ireland, Denmark, India, Poland, Sweden, Argentina, Netherlands, Brazil, Macao, Singapore, Sao Tome and Principe, Cameroon, Hong Kong, Portugal, Morocco, Hungary, Finland, Puerto Rico, Holy See (Vatican City State), Gabon, Bulgaria, Norway, Jamaica
DMEM (Dulbecco's Modified Eagle's Medium) is a cell culture medium formulated to support the growth and maintenance of a variety of cell types, including mammalian cells. It provides essential nutrients, amino acids, vitamins, and other components necessary for cell proliferation and survival in an in vitro environment.
3
Raw264.7 cell by American Type Culture Collection
Sourced in United States, United Kingdom, China, Japan
RAW264.7 cells are a murine macrophage cell line derived from the Abelson murine leukemia virus-induced tumor. They are an adherent cell type that can be used for various cell biology and immunology research applications.
4
Lipopolysaccharides (lps) by Merck Group
Sourced in United States, Germany, China, United Kingdom, Sao Tome and Principe, Macao, Italy, Japan, Canada, France, Switzerland, Israel, Australia, Spain, India, Ireland, Brazil, Poland, Netherlands, Sweden, Denmark, Hungary, Austria, Mongolia
The LPS laboratory equipment is a high-precision device used for various applications in scientific research and laboratory settings. It is designed to accurately measure and monitor specific parameters essential for various experimental procedures. The core function of the LPS is to provide reliable and consistent data collection, ensuring the integrity of research results. No further details or interpretations can be provided while maintaining an unbiased and factual approach.
5
Penicillin streptomycin by Thermo Fisher Scientific
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Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.
6
Trizol reagent by Thermo Fisher Scientific
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TRIzol reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components designed for the isolation of total RNA, DNA, and proteins from a variety of biological samples. The reagent maintains the integrity of the RNA while disrupting cells and dissolving cell components.
7
Streptomycin by Thermo Fisher Scientific
Sourced in United States, United Kingdom, Germany, China, France, Canada, Australia, Japan, Switzerland, Italy, Belgium, Israel, Austria, Spain, Netherlands, Poland, Brazil, Denmark, Argentina, Sweden, New Zealand, Ireland, India, Gabon, Macao, Portugal, Czechia, Singapore, Norway, Thailand, Uruguay, Moldova, Republic of, Finland, Panama
Streptomycin is a broad-spectrum antibiotic used in laboratory settings. It functions as a protein synthesis inhibitor, targeting the 30S subunit of bacterial ribosomes, which plays a crucial role in the translation of genetic information into proteins. Streptomycin is commonly used in microbiological research and applications that require selective inhibition of bacterial growth.
8
Penicillin by Thermo Fisher Scientific
Sourced in United States, United Kingdom, Germany, China, France, Canada, Japan, Australia, Switzerland, Italy, Israel, Belgium, Austria, Spain, Brazil, Netherlands, Gabon, Denmark, Poland, Ireland, New Zealand, Sweden, Argentina, India, Macao, Uruguay, Portugal, Holy See (Vatican City State), Czechia, Singapore, Panama, Thailand, Moldova, Republic of, Finland, Morocco
Penicillin is a type of antibiotic used in laboratory settings. It is a broad-spectrum antimicrobial agent effective against a variety of bacteria. Penicillin functions by disrupting the bacterial cell wall, leading to cell death.
9
Raw 264.7 cells by American Type Culture Collection
Sourced in United States, United Kingdom, Germany, China, Switzerland
RAW 264.7 cells are a mouse macrophage cell line derived from the ascites of a tumor induced by the Abelson murine leukemia virus. The cells are commonly used in research for studying macrophage function and immunology.
10
Raw264 by American Type Culture Collection
Sourced in United States, China, Germany, France, United Kingdom, Italy, Spain, Japan
RAW264.7 is a mouse macrophage cell line derived from the BALB/c mouse. It is a commonly used model for studying macrophage biology and function.

More about "RAW 264.7 Cells"

RAW 264.7 cells are a widely used murine macrophage cell line derived from the ascites of a tumor induced by the Abelson murine leukemia virus.
These versitile cells are a valuable model system for investigating cellular processes and testing therapeutic interventions related to macrophage biology, including inflammation and immune response.
Macrophages are a type of white blood cell that play a crucial role in the immune system, engulfing and digesting cellular debris, pathogens, and other harmful or foreign substances.
RAW 264.7 cells are commonly utilized in cell biology research to study macrophage function, making them a powerful tool for understanding the mechanisms underlying inflammatory and immune-related diseases.
When culturing RAW 264.7 cells, it is important to supplement the growth medium, typically Dulbecco's Modified Eagle's Medium (DMEM), with fetal bovine serum (FBS) and antibiotics such as penicillin and streptomycin to maintain a sterile environment and support cell growth.
Additionally, researchers may employ techniques like TRIzol reagent to isolate and analyze cellular components, such as RNA, for further investigation.
Whether you're studying macrophage biology, testing novel therapeutic interventions, or exploring the cellular processes underlying inflammation and immunity, RAW 264.7 cells offer a robust and well-characterized model system to advance your research.
By leveraging the insights and resources available, you can optimize your experiments and achieve meaningful, reproducible results.