Bone marrow was collected from 12- to 16-week-old female wild-type C57BL/6 mice (experiments approved by the animal experiments committee of the Academic Medical Center, Amsterdam) and cultured in 100 mm cell culture dishes (BD Falcon) in IMDM (Lonza) supplemented with 10% FBS, 86 μg ml−1 gentamicin and 40 ng ml−1 recombinant mouse M-CSF (Biolegend). At day 3, fresh medium containing M-CSF was added. At day 6, M-CSF and either 10 ng ml−1 recombinant mouse IL-10 or 20 ng ml−1 recombinant mouse IL-4 (both from eBioscience) were added. At day 7, cells were harvested and stimulated as described for human macrophages, using 2 μg ml−1 mouse IgG (Equitech-Bio) coated in Maxisorp plates. Cells were analysed by quantitative RT–PCR as described; primer pairs are listed in Supplementary Table 2 .
100 mm cell culture dishes
Manufactured by BD
Sourced in Germany
The 100 mm cell culture dishes are circular, sterile, and disposable containers used for the in vitro cultivation of cells. They provide a consistent, controlled environment for cell growth and experimentation.
Automatically generated - may contain errors
Lab products found in correlation
Gentamicin, by BioLegend (1 mentions)
M csf, by Thermo Fisher Scientific (1 mentions)
Iscove modified dulbecco media (imdm), by Lonza (1 mentions)
Recombinant mouse m csf, by BioLegend (1 mentions)
Recombinant mouse il 10, by Thermo Fisher Scientific (1 mentions)
Recombinant mouse il 4, by Thermo Fisher Scientific (1 mentions)
Gentamicin, by Thermo Fisher Scientific (1 mentions)
75 cm2 cell culture flask, by Corning (1 mentions)
2 protocols using 100 mm cell culture dishes
1
Murine Macrophage Differentiation Protocol
2
Isolation and Expansion of Human MSCs
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Human BM aspirate from healthy donors (ethical approval 11-101-0303, Ethical Committee of the University Hospital of Regensburg) was diluted with DMEM supplemented with 10% FBS and 2.5 mg/ml gentamicin (Life Technologies, Darmstadt, Germany) and plated in 100 mm cell culture dishes (BD Falcon, Heidelberg, Germany). Medium was changed twice weekly and cell growth and morphology were regularly checked and documented using a Hund Willovert S microscope (Hund Wetzlar, Wetzlar, Germany). When confluence was reached, MSCs were trypsinized and further expanded in 75 cm 2 cell culture flasks (Corning Costar, Bodenheim, Germany). After 2-4 weeks (70-90% confluence), MSCs were harvested and phenotypically and functionally analyzed. The specific MSC phenotype was determined by flow cytometric analysis of cell surface proteins as described below.
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