To analyze the differentiation potential of MMPs, single MPP cells were sorted by MoFlo XDP (Beckman Coulter) and plated (one cell/well) in the MEM alpha (Life technology) medium containing 20% ES cell-grade FBS (Millipore), SCF (50 ng/ml: BioLegend), and Flt3 ligand (30 ng/ml: BioLegend) on the layer of OP9 cells in 96-well plates as previously described33 (link). The OP9 cell line was obtained from ATCC and confirmed to be free of mycoplasma by using the MycoAlert PLUS kit (Lonza). Two days after cell plating, IL-3 (10 ng/ml; BioLegend), IL-7 (10 ng/ml; BioLegend), and GM-CSF (10 ng/ml; R&D Systems) were added to each well to induce cell differentiation. “Plating efficiency” denotes cells successfully expanded in culture, determined by microscopic observation. Expanded cells were analyzed by flow cytometry to determine whether they developed into either granulocyte/macrophages (GM) (Gr-1+CD11b+B220−CD19− and Gr-1−CD11b+B220−CD19−) or B cells (Gr-1−CD11b−B220+CD19+).
Es cell grade fbs
Manufactured by Merck Group
ES cell-grade FBS is a high-quality fetal bovine serum (FBS) product designed for use in embryonic stem (ES) cell culture applications. It provides the essential growth factors and nutrients necessary to support the proliferation and maintenance of ES cells in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Interleukin 7 (il 7), by BioLegend (2 mentions)
Mycoalert plus kit, by Lonza (2 mentions)
Flt3 ligand, by BioLegend (2 mentions)
Interleukin 3 (il 3), by BioLegend (2 mentions)
Stem cell factor (scf), by BioLegend (2 mentions)
Moflo xdp, by Beckman Coulter (2 mentions)
Alpha mem, by Thermo Fisher Scientific (2 mentions)
Gm csf, by R&D Systems (2 mentions)
2 protocols using es cell grade fbs
1
Differentiation potential of single MPP cells
2
Differentiation potential of single MPP cells
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To analyze the differentiation potential of MMPs, single MPP cells were sorted by MoFlo XDP (Beckman Coulter) and plated (one cell/well) in the MEM alpha (Life technology) medium containing 20% ES cell-grade FBS (Millipore), SCF (50 ng/ml: BioLegend), and Flt3 ligand (30 ng/ml: BioLegend) on the layer of OP9 cells in 96-well plates as previously described33 (link). The OP9 cell line was obtained from ATCC and confirmed to be free of mycoplasma by using the MycoAlert PLUS kit (Lonza). Two days after cell plating, IL-3 (10 ng/ml; BioLegend), IL-7 (10 ng/ml; BioLegend), and GM-CSF (10 ng/ml; R&D Systems) were added to each well to induce cell differentiation. “Plating efficiency” denotes cells successfully expanded in culture, determined by microscopic observation. Expanded cells were analyzed by flow cytometry to determine whether they developed into either granulocyte/macrophages (GM) (Gr-1+CD11b+B220−CD19− and Gr-1−CD11b+B220−CD19−) or B cells (Gr-1−CD11b−B220+CD19+).
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