Raw 264

Manufactured by RIKEN BioResource Center

Sourced in Japan

RAW 264.7 is a mouse macrophage cell line. It is widely used in cell biology research. The cells can be cultured in vitro and used for various experimental applications.

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Lab products found in correlation

Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (10 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (9 mentions) Fetal bovine serum (fbs), by Merck Group (4 mentions) Dulbecco modified eagle medium (dmem), by Fujifilm (4 mentions) Raw264, by Thermo Fisher Scientific (4 mentions) Penicillin streptomycin, by Merck Group (3 mentions) Lipopolysaccharide lps, by Merck Group (3 mentions) Perfect real time supporting system, by Takara Bio (2 mentions) Lipopolysaccharides (lps), by Merck Group (2 mentions) Lps from e coli o111 b4, by Merck Group (2 mentions) Poly l lysine (pll), by Merck Group (2 mentions) Pll solution, by Merck Group (2 mentions) Dulbecco modified eagle medium (dmem), by Nacalai Tesque (2 mentions) Raw264, by Nacalai Tesque (2 mentions) L glutamine, by Merck Group (2 mentions) Dulbecco s modified eagle medium, by Thermo Fisher Scientific (2 mentions) Penicillin, by Sartorius (2 mentions) Streptomycin, by Sartorius (2 mentions) Minimum essential medium (mem), by Nacalai Tesque (1 mentions) Mc3t3 e1, by Nacalai Tesque (1 mentions)

Close spelling variants of this product

RAW 264.7 macrophages (5 citations) RAW 264.7 murine macrophage cell line (5 citations) RAW 264.7 (BCRC 60001) (2 citations)

47 protocols using raw 264

Mouse Cell Line Culturing Protocol

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A mouse skeletal myoblast
(C2C12) cell line (RCB0987) and mouse leukemic monocyte (RAW264) cell
line (RCB0535) were purchased from the RIKEN BRC Cell Bank (Ibaraki,
Japan). The plastic dishes (Falcon 353002) were purchased from Corning
(Corning, NY). The 96-well plates (92096) were purchased from TRP
(Trasadingen, Switzerland). The glass capillaries with a polyimide
coating (inner diameter (id) of 40, 180, or 250 μm, outer diameter
(od) of 100 or 360 μm) were purchased from Molex (Lisle, IL).

Ota N., Tanaka N., Sato A., Shen Y., Yalikun Y, & Tanaka Y. (2022). Microenvironmental Analysis and Control for Local Cells under Confluent Conditions via a Capillary-Based Microfluidic Device. Analytical Chemistry, 94(47), 16299-16307.

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Culturing Murine Macrophages and Osteoblasts

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The murine macrophage-like cell line RAW264 was purchased from Riken BRC (Ibaraki, Japan). RAW264 cells were cultured in a minimum essential medium (MEM; Nacalai Tesque) with 10% fetal bovine serum (FBS; Sigma), nonessential amino acid solution (×100; Nacalai Tesque), and penicillin-streptomycin-amphotericin B suspension (×100; FUJIFILM Wako, Osaka, Japan). Murine calvaria-derived osteoblast-like cell line (OBs) MC3T3-E1 was also purchased from Riken BRC. MC3T3-E1 cells were cultured in a medium containing αMEM (Nacalai Tesque) supplemented with 10% FBS and penicillin-streptomycin-amphotericin B suspension. Both cells were passaged twice a week and maintained at 37 °C and 5% CO₂.

Ueda K., Ma C., Izumiya M., Kuroda C., Ishida H., Uemura T., Saito N., Aoki K, & Haniu H. (2023). Biocompatibility Evaluation of Carbon Nanohorns in Bone Tissues. Nanomaterials, 13(2), 244.

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LPS-Induced Cytokine Expression in RAW264 Cells

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Mouse monocyte/macrophage cell line RAW264 was obtained from RIKEN BRC
(Ibaraki, Japan). The cells were grown in Dulbecco’s modified Eagle
medium (DMEM: Wako) containing 10% fetal bovine serum with 100 U/ml of
penicillin G and 100 μg/ml of streptomycin at 37°C in 5%
CO₂ with a humidified atmosphere. Subconfluent cells in 12-well
plates were exposed to LPS (Sigma-Aldrich) at a concentration of 1 μg/ml,
and total RNA was extracted 4 h later. Subsequently, reverse transcription and
PCR were performed as described in the previous section. Predesigned primer sets
for mouse TNF-α (primer set MA097070), IL-6 (primer set MA039013), IL-12A
(primer set MA0287533), IL-23A (primer set MA095159), and TBP (primer set
MA050367) were obtained from the Perfect Real-Time Supporting System (Takara
Bio). The messenger RNA expression level was normalized to the expression of
TBP. CN210 (0.1, 1 and 10 μM) and dexamethasone (1 μM) were
cotreated with LPS in RAW264 cells.

Noguchi T., Hidaka K., Kobayashi S., Matsumoto K., Yoshioka M., Hu X., Maloney D.J., Yang S.M, & Kato S. (2021). A quinazoline-based bromodomain inhibitor, CN210, ameliorates indomethacin-induced ileitis in mice by inhibiting inflammatory cytokine expression. Drug development research, 82(8), 1235-1246.

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Culturing Mouse Macrophage-like RAW264 Cells

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The mouse macrophage-like cell line RCB053, named RAW264 (RIKEN Bio Resource Center, Tsukuba, Japan), was maintained in a minimum essential medium (MEM, Thermo Fisher Scientific) containing 2 mM l-glutamine, 10% (v/v) fetal bovine serum (FBS) (Sigma-Aldrich, St Louis, MO, USA), and 1% (v/v) MEM non-essential amino acid solution (100×; Fujifilm Wako Pure Chemical, Osaka, Japan). The cells were maintained in a humidified incubator with 5% CO₂ at 37 °C. For the assay, penicillin and streptomycin mixtures (Thermo Fisher Scientific) were added to the cell culture medium to final concentrations of 100 units/mL and 100 µg/mL, respectively.

Zhao D., Tu A.T., Shobo M., Le N.B., Yoshikawa C., Sugai K., Hakamata Y, & Yamazaki T. (2022). Non-Modified CpG Oligodeoxynucleotide Forming Guanine-Quadruplex Structure Complexes with ε-Poly-L-Lysine Induce Antibody Production as Vaccine Adjuvants. Biomolecules, 12(12), 1868.

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Murine and Human Macrophage Culture

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Murine macrophage cells, RAW 264 (RIKEN BRC CELL bank, Ibaraki, Japan), were incubated in Dulbecco’s Modified Eagle’s Medium (DMEM) high glucose (Sigma-Aldrich, St Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich), 50 U mL⁻¹ penicillin, and 50 μg mL⁻¹ streptomycin. U937 cells (JCRB Cell Bank, Osaka, Japan), a human monocyte cell line, were cultured in RPMI-1640 (Sigma-Aldrich, St Louis, MO, USA) supplemented with 10% FBS (Sigma-Aldrich). The cells were incubated at 37 °C in a humidified atmosphere containing 5% CO₂.

Nakamura Y., Matsumoto H., Wu C.H., Fukaya D., Uni R., Hirakawa Y., Katagiri M., Yamada S., Ko T., Nomura S., Wada Y., Komuro I., Nangaku M., Inagi R, & Inoue T. (2023). Alpha 7 nicotinic acetylcholine receptors signaling boosts cell-cell interactions in macrophages effecting anti-inflammatory and organ protection. Communications Biology, 6, 666.

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Culturing and Stimulating Raw264 Macrophages

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Raw264 (Riken BioResource Center) are cultured in Dulbecco's modified Eagle medium (DMEM, Nacalai) supplemented with 10% fetal bovine serum (Gibco) and penicillin/streptomycin (Sigma-Aldrich) with 10 000 units and 10 mg mL -1 diluted at 10 mL L -1 , respectively. Cells are plated on 10 cm tissueculture dishes and incubated at 37 °C in a humidified atmosphere with 5% CO 2 . For observation in the Raman system, cells are first detached from the dish with a solution containing 0.25% trypsin and 1 mM ethylenediaminetetraacetic acid (Nacalai) for approximately 5 minutes at 37 °C. The cell suspension is then plated at a density of 30 000 cells per cm 2 on quartz dishes (FPI) pre-coated with poly-L-lysine (PLL, Sigma-Aldrich) by immersing the surface in a 0.01% PLL solution (Sigma-Aldrich) for 30 min at room temperature (RT). Cells are then incubated for 5-6 hours to allow them to adhere to the dish substrate. They are then stimulated by replacing the culture medium with fresh DMEM containing lipopolysaccharide (LPS) from E. coli O111:B4 (Sigma-Aldrich) and/or cycloheximide (CHX, Sigma-Aldrich). Cells are then incubated for 20-21 hours before measurements.

Pavillon N, & Smith N.I. (2021). Deriving accurate molecular indicators of protein synthesis through Raman-based sparse classification. The Analyst, 146(11).

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Culturing Murine Macrophage-like RAW 264 Cells

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The murine macrophage-like cell line, RAW 264 (RCB0535), was provided from the RIKEN BioResource Center (Ibaraki, Japan). RAW 264 cells were cultured in DMEM containing 10% FBS, 100 units/mL penicillin, and 100 μg/ mL streptomycin. Cells were incubated at 37 °C with 5% CO 2 in a humidified atmosphere.

Azuma S., Murakami Y., Azuma E., Baba K., & Taniguchi M. (2020). Anti-inflammatory effects of flavonoids in Citrus jabara fruit peels. Trends in Immunotherapy, 4(1), 5-5.

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Raman Spectroscopy of Stimulated RAW264 Cells

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Cell culture and stimulation Raw264 (Riken BioResource Center) are cultured in Dulbecco's modified Eagle medium (DMEM, Nacalai) supplemented with 10% fetal bovine serum (Gibco) and penicillin/streptomycin (Sigma-Aldrich) with 10,000 units and 10 mg/mL diluted at 10 mL/L, respectively. Cells are plated on 10-cm tissue-culture dishes and incubated at 37 • C in a humidified atmosphere with 5% CO 2 . For observation in the Raman system, cells are first detached from the dish with a solution containing 0.25% trypsin and 1 mM ethylenediaminetetraacetic acid (Nacalai) for approximately 5 minutes at 37 • C. The cell suspension is then plated at a density of 30, 000 cells/cm 2 on quartz dishes (FPI) pre-coated with poly-L-lysine (PLL, 2 Sigma-Aldrich) by immersing the surface in a 0.01% PLL solution (Sigma-Aldrich) for 30 min at room temperature (RT). Cells are then incubated for 5-6 hours to allow them to adhere to the dish substrate. They are then stimulated by replacing the culture medium with fresh DMEM containing lipopolysaccharide (LPS) from E. Coli O111:B4 (Sigma-Aldrich) and/or cycloheximide (CHX, Sigma-Aldrich). Cells are then incubated for 20-21 hours before measurements.

Pavillon N., & Smith N.I. (2021). Deriving accurate molecular indicators of protein synthesis through Raman-based sparse classification.

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Evaluating Anti-inflammatory Effects of Compounds on RAW264 Cells

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Mouse monocyte/macrophage cell line RAW264 were obtained from RIKEN BRC (Ibaraki, Japan). The cells were grown in Dulbecco's modified Eagle medium (DMEM: Wako) containing 10% fetal bovine serum (FBS) with 100 U/mL of penicillin G and 100 µg/mL of streptomycin at 37 ˚C in 5% CO 2 with a humidified atmosphere. Sub-confluent cells in 12-well plate were exposed of lipopolysaccharide (LPS) (Sigma-Aldrich) at a concentration of 1 µg/mL and total RNA was extracted 4 h later. Subsequently reverse transcription and PCR were performed as described above. Pre-designed primer sets for mouse TNF-α (primer set MA097070), IL-6 (primer set MA039013), IL-12A (MA0287533), IL-23A (MA095159), and TATA-binding protein (TBP) (MA050367) were obtained from the Perfect Real-Time Supporting System (Takara Bio). The mRNA expression level was standardised to that of TBP.
. CC-BY 4.0 International license perpetuity. It is made available under a preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in The copyright holder for this this version posted January 24, 2020. ; https://doi.org/10.1101/2020.01.24.917948 doi: bioRxiv preprint CN210 at 0.1, 1 and 10 μM and dexamethasone at 1 μM were co-treated with LPS in RAW264 cells.

Noguchi T., Hidaka K., Kobayashi S., Matsumoto K., Yoshioka M., Yang S., Maloney D.J., & Kato S. (2020). Ameliorative effect of a quinazoline-based bromodomain inhibitor, CN210, on experimentally-induced Crohn’s disease-like murine ileitis via inhibition of inflammatory cytokine expression.

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Mouse 3T3-L1 Preadipocyte Differentiation

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Mouse 3T3-L1 preadipocytes (American Type Culture Collection, Manassas, VA) were cultured in Dulbecco's modified Eagle's medium supplemented with 100 units/mL of penicillin G and 0.1 mg/mL of streptomycin (DMEM) containing 10% calf serum. Differentiation of 3T3-L1 preadipocytes into adipocytes was achieved by the following procedure: cells were cultured in the initiation medium of DMEM containing 10% fetal calf serum (FCS), 5 µg/mL insulin (Sigma-Aldrich), 0.5 mM 3-isobutyl-1-methylxanthine (Wako Pure Chemical, Osaka, Japan), and 0.25 µM dexamethasone (Sigma-Aldrich) for 2 days. After 3 days of culture in the progression medium (DMEM containing 10% FCS and 5 µg/mL insulin), cells were cultured in the maintenance medium (DMEM containing 10% FCS), which was exchanged every other day. Cells were used for experiments at days 10-14 after the induction of differentiation. RAW264 (RIKEN BioResource Center, Tsukuba, Japan) and J774A.1 (American Type Culture Collection) macrophage-like cell lines were cultured in DMEM containing 10% FCS. Mouse peritoneal exudate cells were prepared as described previously (Schneider, 2013) .

Furuoka M., Ozaki K., Sadatomi D., Mamiya S., Yonezawa T., Tanimura S, & Takeda K. (2016). TNF-α Induces Caspase-1 Activation Independently of Simultaneously Induced NLRP3 in 3T3-L1 Cells. Journal of cellular physiology, 231(12).

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