A mouse macrophage cell line (RAW264.7) was obtained from the American Type Culture Collection (Manassas, USA). The RAW264.7 cells were grown in Dulbecco's modified Eagle's medium (DMEM; Nacalai tesque, catalog 08456-36) containing 10% fetal bovine serum (FBS) in a humidified atmosphere of 95% air and 5% CO2 at 37⁰ C. The cells were passaged by trypsinization every 2 to 3 days, and subconfluent monolayers of RAW264.7 cells from passages 10 to 16 were used in the experiments.
Raw264
Manufactured by Nacalai Tesque
Sourced in Japan
RAW264.7 is a mouse macrophage cell line derived from ascites of a tumor induced by the Abelson murine leukemia virus. It is commonly used in cell culture studies for various applications.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Nacalai Tesque (4 mentions)
Raw264, by American Type Culture Collection (3 mentions)
Rpmi 1640 medium, by Nacalai Tesque (2 mentions)
Fetal bovine serum (fbs), by Biowest (2 mentions)
Raw264, by RIKEN Cell Bank (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Minimum essential medium (mem), by Nacalai Tesque (2 mentions)
Raw 264, by RIKEN BioResource Center (2 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Phorbol myristate acetate (pma), by Fujifilm (1 mentions)
Dulbecco s modified eagle medium, by Nacalai Tesque (1 mentions)
Thp 1 cells rcb 1189, by RIKEN Cell Bank (1 mentions)
Human recombinant soluble rankl, by Oriental Yeast (1 mentions)
Raw 264, by R&D Systems (1 mentions)
Tib 71, by American Type Culture Collection (1 mentions)
Penicillin streptomycin mixed solution, by Nacalai Tesque (1 mentions)
Dulbecco s modified eagle, by Nacalai Tesque (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Nacalai Tesque (1 mentions)
10 protocols using raw264
1
Culturing RAW264.7 Mouse Macrophages
2
Immortalized Human DP-1 Cell Line Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The establishment of immortalized human DP-1 cells has been described previously [9 (link)]. Human monocytic leukemia THP-1 cells (RCB1189) and the mouse macrophage cell line RAW264.7 were purchased from RIKEN Cell Bank (Tsukuba, Japan) and American Type Culture Collection (ATCC, Manassas, VA), respectively. DP-1 and RAW264.7 cells were cultured in Dulbecco's Modified Eagle Medium (Nacalai Tesque, Kyoto, Japan) supplemented with 10% fetal bovine serum (FBS) (Biowest, France), 100 U/ml penicillin, and 100 μg/ml streptomycin. THP-1 cells were maintained in RPMI 1640 medium (Nacalai Tesque) containing 10% FBS, 2 mM GlutaMAX™ (Gibco, Thermo Fisher Scientific, Waltham, MA), and antibiotics.
THP-1 cells were seeded in 6-well plates at a density of 5 × 105 cells/well and stimulated with 100 nM PMA (FUJIFILM Wako Pure Chemical, Osaka, Japan) for 24 h to obtain PMA-differentiated macrophages.
THP-1 cells were seeded in 6-well plates at a density of 5 × 105 cells/well and stimulated with 100 nM PMA (FUJIFILM Wako Pure Chemical, Osaka, Japan) for 24 h to obtain PMA-differentiated macrophages.
3
Murine Macrophage and Rat Stem Cell Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The murine macrophage cell line RAW264.7 (EC91062702; KAC Co., Kyoto, Japan) was used to evaluate the inflammatory response to the samples in our research. RAW264.7 cells were maintained in high-glucose Dulbecco’s modified Eagle’s medium (DMEM; Nacalai Tesque, Inc., Kyoto, Japan) supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich, St. Louis, MO, USA) and 1% penicillin/streptomycin. Moreover, the cells were cultured at 37 °C with 5% CO2. Adherent cells were dislodged by gently passing a cell scraper upon reaching approximately 80% confluence.
Rat bone marrow mesenchymal stem cells (rBMMSCs) were used to evaluate the osseointegration of the samples. Rat BMMSCs were isolated from the femurs of 8-week-old Sprague–Dawley rats (Shimizu Laboratory Supplies Co., Kyoto, Japan) and maintained in minimum essential medium (MEM, Nacalai Tesque, Inc., Kyoto, Japan) with 10% FBS and 1% penicillin/streptomycin [86 (link)]. The medium was replaced every third day. The confluent cells were subcultured by trypsinization, and cells at passages 3–5 were used for subsequent studies.
Rat bone marrow mesenchymal stem cells (rBMMSCs) were used to evaluate the osseointegration of the samples. Rat BMMSCs were isolated from the femurs of 8-week-old Sprague–Dawley rats (Shimizu Laboratory Supplies Co., Kyoto, Japan) and maintained in minimum essential medium (MEM, Nacalai Tesque, Inc., Kyoto, Japan) with 10% FBS and 1% penicillin/streptomycin [86 (link)]. The medium was replaced every third day. The confluent cells were subcultured by trypsinization, and cells at passages 3–5 were used for subsequent studies.
4
Osteoclast Differentiation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mouse osteoclast precursor cell line RAW 264.7 (RCB0535) from the Riken BioResource Center (BRC) was co-cultured with the supernatants from the L929 cells cultured on each substrate. The cells were grown in the RAW 264.7 medium consisting of the minimum essential medium and alpha modification (Nacalai Tesque, Kyoto, Japan) supplemented with 10% FBS at 37 °C in a humidified 5% CO2 atmosphere. The cells were then passaged twice. At 70% confluency, the cells were detached by scraping and seeded on 24- or 96-well culture-graded polystyrene plates at 7 × 104 cells/mL or 1.125 × 104 cells/mL in the RAW 264.7 medium. According to a previously reported protocol, 50 ng/mL recombinant human soluble RANKL (Oriental Yeast, Tokyo, Japan) and 50 ng/mL TNF-α (aa80-235, R&D System, Minneapolis, MN, USA) were added to the RAW 264.7 culture medium for osteoclast differentiation. Fresh DMEM/F12 basal medium or the L929 culture supernatants were mixed with RAW 264.7 medium in a 1:9 volume ratio for 5 days and then evaluated for TRAP staining, osteoclast cell number, and osteoclast-related marker gene expression.
5
Culturing Murine Macrophages and Human PBMCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A murine macrophage cell line RAW264.7 (TIB-71™) was obtained from American Type Culture Collection (ATCC; Manassas, VA, USA). RAW264.7 cells were cultured in Dulbecco’s modified Eagle’s medium with high glucose (Nacalai Tesque) containing 10% heat-inactivated FBS (Biowest, Vieux Bourg, France) and a penicillin-streptomycin Mixed Solution (Nacalai Tesque). Human CD14+ PBMCs were purchased from Lonza (Basel, Switzerland) and cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Nacalai Tesque) containing 10% heat-inactivated fetal bovine serum (FBS). In this study, no human studies were conducted.
6
Culturing Murine Preosteoblasts and Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Murine calvaria-derived preosteoblastic cell line MC3T3-E1 subclone 4 and murine macrophage cell line RAW264.7 were obtained from ATCC (Manassas, VA, USA). MC3T3-E1 cells, primary osteoblasts, and primary osteoclast precursors were cultured in α-MEM with 10% FBS and 1% penicillin-streptomycin at 37°C with 5% CO2. RAW264.7 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Nacalai Tesque) with 10% FBS and 1% penicillin-streptomycin at 37°C with 5% CO2.
7
Culturing Mouse Macrophage RAW264.7 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A mouse macrophage cell line (RAW264.7) was obtained from the American Type Culture Collection (Manassas, USA). The RAW264.7 cells were grown in Dulbecco’s modified Eagle’s medium (DMEM; Nacalai Tesque, catalog 08456-36) containing 10% fetal bovine serum (FBS) in a humidified atmosphere of 95% air and 5% CO2 at 37 °C. The cells were passaged by trypsinization every 2 to 3 days, and subconfluent monolayers of RAW264.7 cells from passages 10 to 16 were used in the experiments.
8
Culturing Murine Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The murine macrophage-like cell line RAW264 and the murine colon adenocarcinoma cell line colon-26 were purchased from RIKEN Cell Bank (Tsukuba, Japan). RAW264 cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Nacalai Tesque, Kyoto, Japan) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin and 100 µg/mL streptomycin (Gibco, Waltham, MA, USA). colon-26 cells were cultured in RPMI1640 (Nacalai Tesque) supplemented with 10% FBS, 100 U/mL penicillin, and 100 µg/mL streptomycin. These cell lines were cultured at 37 °C in a 5% CO2 incubator.
9
Osteoclast Formation Assay with RAW 264 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mouse leukemic monocyte–macrophage cell line RAW 264 was purchased from the Japan Riken BioResource Research Center (RCB0535) (Ibaraki, Japan). RAW264 cells (1 × 103 cells/well) were seeded into 96-well plates and cultured in α-minimal essential medium (Nacalai Tesque Inc., Kyoto, Japan) containing 10% fetal bovine serum (HyClone, Logan, UT, USA) and 100 units/mL penicillin–streptomycin (Thermo Fisher Scientific, Waltham, MA, USA). Kampo medicines at the concentration of 300 µg/mL in 0.1% DMSO or 0.1% DMSO (control) were added to the wells. Cells were incubated at 37 °C in a 5% CO2 atmosphere for three to five days with 100 ng/mL sRANKL to induce osteoclast formation. Osteoclasts were confirmed by TRAP staining. TRAP-positive mononuclear (nuclei/cell = 1) and multinucleated osteoclasts (nuclei/cell ≥ 3) were counted under a microscope.
10
Culturing Murine Macrophages and Osteoblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!