Sphere formation was determined as previously described [20 (link)]. Cells were seeded on the ultra–low attachment 24-well plates (Corning Costar) with sphere formation medium, which was contained serum free culture medium, B27 supplement, recombinant human EGF (20 ng/ml), and basic FGF (10 ng/ml) for 14 days. The diameters of spheres reached 50 μm were counted.
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Sphere Formation Assay Protocol
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Corresponding Organization : National Taiwan University Hospital
Other organizations : National Defense Medical Center, Tri-Service General Hospital, Development Center for Biotechnology
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Variable analysis
independent variables
- Seeding cells on ultra–low attachment 24-well plates
- Diameters of spheres reaching 50 μm
- Sphere formation medium, which contained serum free culture medium, B27 supplement, recombinant human EGF (20 ng/ml), and basic FGF (10 ng/ml)
- No positive or negative controls were explicitly mentioned.
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