Bone marrow was flushed from the tibiae and femurs of 8–10-week-old male C57BL/6J mice using a 25-gauge needle and DMEM media (ATCC) containing 10% FBS, 20 mM HEPES and 1% penicillin/streptomycin. Once isolated, the bone marrow was plated in this medium supplemented with 10 ng/ml M-CSF (macrophage colony-stimulating factor; Sigma). On day four of differentiation, the media was replenished with fresh media (containing M-CSF). After seven days, the cells were liberated using a cell scraper centrifuged and re-plated in media lacking M-CSF. After 24 hours, cells were exposed to 10 nM RvD2 (Cayman Chemical) for four hours then collected in Buffer RLT (Qiagen) with 10 μl/ml 2-mercaptoethanol.
Dmem media
Manufactured by American Type Culture Collection
Sourced in United States
DMEM media is a commonly used cell culture medium formulation developed by the American Type Culture Collection (ATCC). It is a powdered, basal medium that provides essential nutrients for the growth and maintenance of a variety of cell types in in vitro cell culture systems. The core function of DMEM media is to support the basic nutritional requirements of cells.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by American Type Culture Collection (3 mentions)
Rpmi 1640 media, by American Type Culture Collection (3 mentions)
Rlt buffer, by Qiagen (2 mentions)
M csf, by Merck Group (2 mentions)
Rpmi 1640 media, by Thermo Fisher Scientific (2 mentions)
Rpmi8226, by American Type Culture Collection (2 mentions)
Dmem media, by Thermo Fisher Scientific (2 mentions)
Nci h929, by American Type Culture Collection (2 mentions)
Hepa1 6, by American Type Culture Collection (2 mentions)
Penicillin streptomycin, by Corning (2 mentions)
Mia paca 2, by American Type Culture Collection (2 mentions)
Bxpc 3, by American Type Culture Collection (2 mentions)
Panc 1, by American Type Culture Collection (2 mentions)
Aspc 1, by American Type Culture Collection (2 mentions)
Rpmi 1640, by American Type Culture Collection (2 mentions)
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
Trypan blue, by Merck Group (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
19 protocols using dmem media
1
Isolating Mouse Bone Marrow Macrophages
2
Cell Culture Conditions for Myeloma and CLL
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Cells were cultured in incubators containing
5% CO2 and maintained at 37 °C. RPMI-8226 and NCI-H929
human MM cell lines (purchased from ATCC), 5TGM1 mouse MM cell line
(a gift from Dr. Lori A. Hazlehurst at the West Virginia University,
Morgantown, WV), MEC2 and WaC3 human CLL cell lines (gifts from Dr.
Javier A. Pinilla-Ibarz at the Moffitt Cancer Center, Tampa, FL),
and primary B cells purified from spleens of mice were grown in RPMI
1640 media (Gibco) supplemented with heat-inactivated fetal bovine
serum (FBS, 10%),l -glutamine (2 mM), sodium pyruvate (1
mM), nonessential amino acids (0.1 mM), β-mercaptoethanol (β-ME;
0.1 mM), penicillin G sodium (100 U/mL), and streptomycin sulfate
(100 μg/mL). The J558 mouse myeloma cell line (purchased from
ATCC) was cultured in DMEM media (Gibco) and 10% heat-inactivated
horse serum together with the abovementioned supplemental nutrients.
Human embryonic kidney 293 T cells (purchased from ATCC) and mouse
hepatoma HEPA 1–6 cell line (purchased from ATCC) were cultured
in DMEM media with 10% heat-inactivated FBS and the same supplemental
nutrients.
5% CO2 and maintained at 37 °C. RPMI-8226 and NCI-H929
human MM cell lines (purchased from ATCC), 5TGM1 mouse MM cell line
(a gift from Dr. Lori A. Hazlehurst at the West Virginia University,
Morgantown, WV), MEC2 and WaC3 human CLL cell lines (gifts from Dr.
Javier A. Pinilla-Ibarz at the Moffitt Cancer Center, Tampa, FL),
and primary B cells purified from spleens of mice were grown in RPMI
1640 media (Gibco) supplemented with heat-inactivated fetal bovine
serum (FBS, 10%),
mM), nonessential amino acids (0.1 mM), β-mercaptoethanol (β-ME;
0.1 mM), penicillin G sodium (100 U/mL), and streptomycin sulfate
(100 μg/mL). The J558 mouse myeloma cell line (purchased from
ATCC) was cultured in DMEM media (Gibco) and 10% heat-inactivated
horse serum together with the abovementioned supplemental nutrients.
Human embryonic kidney 293 T cells (purchased from ATCC) and mouse
hepatoma HEPA 1–6 cell line (purchased from ATCC) were cultured
in DMEM media with 10% heat-inactivated FBS and the same supplemental
nutrients.
3
Isolation and Characterization of Alveolar Epithelial Type II Cells
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AEC2s were isolated from normal mice using previously described method61 (link) with the following modifications: lungs were subjected to dispase I treatment and cells were passed through 100 and 40 μm filters. Red blood cells were lysed using red blood cells lysis buffer (Sigma–Aldrich, St. Louis, MO, USA; no. R7757). AEC2s were grown on fibronectin-coated (10 μg/ml) plates and cultured in DMEM media (ATCC; no. 30-2002) supplemented with 10% fetal bovine serum and 5% mixture of penicillin G, streptomycin, amphotericin B (Invitrogen) and plasmocin (Invivogen). Purity of AEC2s was measured by considering E-cadherin-positive cells as epithelial cells and vimentin-positive cells as non-epithelial cells.
4
Isolating Mouse Bone Marrow Macrophages
5
Cell Culture of MCF7, HeLa, and HUVEC
6
Culturing Pancreatic Cancer Cell Lines
7
Isolation and Culture of Mouse Macrophages
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To examine phagocytosis in mouse macrophages, bone marrow‐derived macrophages were generated from adult WT and histamine receptor‐deficient C57BL/6J mice by culturing bone marrow cells as previously described (Trouplin et al., 2013 ). Briefly, bone marrow cells were harvested by flushing tibias and femurs, in the presence of 50 nM recombinant murine macrophage colony‐stimulating factor (M‐CSF) (PeproTech, 315–02) in complete DMEM (DMEM media (ATCC, 30‐2002) with heat‐inactivated fetal bovine serum (FBS) (Thermo Fisher, 10438026), and 1 × antibiotic/antimycotic (Thermo Fisher, 15240062)) for seven days. Peritoneal macrophages were elicited from adult mice by intraperitoneal injection of 2% Bio‐Gel P‐100 (Bio‐Rad, 1504174), followed by a 4‐day rest period as previously described (Ray & Dittel, 2010 ). After the rest period, macrophages were harvested by peritoneal lavage with 10 ml prewarmed PBS. The resulting peritoneal cell suspension was washed by centrifugation at 600 g for 10 min at 4˚C and then resuspended in complete DMEM and allowed to attach to nontissue culture treated, plastic Petri dishes for 6 hr before washing twice with PBS and detaching with 0.25% Trypsin‐EDTA (Thermo Fisher, 25200‐056).
8
Cell Viability Assay with Common Lab Reagents
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Verapamil (VRP), and Trypan Blue were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Sulforhodamine-B (SRB) was purchased from Biotium Inc. (Hayward, CA, USA). Penicillin streptomycin and trypsin were purchased from Gibco (Grand Island, NY, USA). Phosphate buffer saline (PBS) was purchased from Becton Dickinson (Fullerton, CA, USA). RPMI-1640 media, DMEM media, fetal bovine serum (FBS), and other cell culture materials were purchased from ATCC (Houston, TX, USA). Other reagents were of the highest analytical grade.
9
Cell Culture Conditions and Maintenance
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All cells were grown in a standard water-jacketed incubator with 5% CO2. K562 cells were grown in IMDM media (#30–2005, ATCC, Manassas, VA) with 10% FBS (#30–2020, ATCC, Manassas, VA) and 1x penicillin/streptomycin. Cells were maintained below 1 million cells per milliliter. HEK293T cells were grown in DMEM media (#30–2002, ATCC, Manassas, VA) with 10% FBS and 1x penicillin/streptomycin. Normocin (1:500) was used as a common additive. All cells were passaged less than 25 times. For passaging, cells were trypsinized with Trypsin-EDTA (#25300–054, Thermo Fisher Scientific, Waltham, MA). Puromycin (2 μg/ml), blasticidin (5 μg/ml), and zeocin (50 μg/ml) were added when necessary for selection. Hct116 cells were grown in McCoy’s 5a modified media (#30–2007, ATCC, Manassas, VA) with 10% FBS (#30–2020, ATCC, Manassas, VA) and 1x penicillin/streptomycin.
10
Culturing Cell Lines for Biomedical Research
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All the cell lines were obtained from ATCC. A549, a human alveolar basal epithelial cell carcinoma, was cultured in phenol red free F-12K medium (ATCC), and Panc-1, a human pancreatic adenocarcinoma cell line, and 293T/17 cells were cultured in phenol red free Dulbecco's Modified Eagle's Medium (DMEM) media (ATCC). The media were supplemented by 10% FBS (Atlanta Biologicals) and 1% penicillin (100 U/mL) /streptomycin (100 μg/mL) solution (Gibco). Murine mesenchymal stem cells (mMSC) were grown in DMEM containing 10% FBS, 10% horse serum (Atlanta Biologicals) and 1% penicillin/streptomycin. Cultures were maintained in a humidified incubator at 37 °C and 5% CO2.
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