Sphere formation assays were performed as reported previously [20 (link)]. MB49 cells were maintained in serum-free CSC culture medium: RPMI supplemented with 20 ng/mL mouse recombinant epidermal growth factor (mEGF) (PeproTech, USA), 20 ng/mL mouse recombinant basic fibroblast growth factor (mbFGF) (PeproTech, USA), 20 ng/mL mouse leukemia inhibitory factor (mLIF) (R&D, MN, USA), B-27 (Gibco, USA), and 0.4% BSA (Sigma, USA). An S2 single-cell suspension maintained in CSC medium was subsequently cultured in ultra-low attachment 6-well plates (Thermo Fisher, USA) at a density of < 5,000 cells/well for 10 days. Spheres that had formed during this period were recorded and colonies with a diameter greater than 50 μm were counted and photographed with a phase-contrast fluorescence microscope (Nikon, ECLIPSE 80i).
Mouse recombinant epidermal growth factor megf
Manufactured by Thermo Fisher Scientific
Sourced in United States
Mouse recombinant epidermal growth factor (mEGF) is a protein that stimulates the growth and differentiation of various cell types. It is a well-characterized growth factor that plays a key role in cellular processes such as proliferation, migration, and survival.
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2 protocols using mouse recombinant epidermal growth factor megf
1
Sphere Formation Assay for Cancer Stem Cells
2
Sphere Formation Assay for Cancer Stem Cells
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Sphere formation assays were performed as reported previously (20) . MB49 cells were maintained in serum-free CSC culture medium: RPMI supplemented with 20 ng/mL mouse recombinant epidermal growth factor (mEGF) (PeproTech, USA), 20 ng/mL mouse recombinant basic broblast growth factor (mbFGF) (PeproTech, USA), 20 ng/mL mouse leukemia inhibitory factor (mLIF) (R&D, MN, USA), B-27 (Gibco, USA), and 0.4% BSA (Sigma, USA). Single S2 cell suspension maintained in CSC medium was subsequently cultured in ultra-low attachment 6-well plates (Thermo Fisher, USA) at a density of < 5,000 cells/well for 10 days. Spheres that had formed during this period were recorded under a microscope and were photographed with a phase-contrast uorescence microscope (Nikon, ECLIPSE 80i).
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