A predefined number of cells was seeded in suspension culture plates (Greiner Bio-One, Kremsmünster, Austria) 24 h after transfection. Cells were then cultivated with Spheromax CSC Medium (Promocell, Heidelberg, Germany). After 7 days, spheres were counted (number per microscopic field of view, 50× magnification) and the area was measured using an open source ImageJ macro, manually correcting visually incorrect counts [65 (link)].
Spheromax csc medium
Manufactured by PromoCell
Sourced in Germany
Spheromax CSC Medium is a cell culture medium specifically designed for the cultivation of cancer stem cells (CSCs). It provides a defined and optimized composition to support the growth and maintenance of CSCs in vitro.
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Lab products found in correlation
3 protocols using spheromax csc medium
1
Sphere Formation Assay Protocol
2
Sphere Formation Assay for MSI-1 Knockdown
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Equal numbers of control and MSI-1 knockdown cells were seeded into coated six-well plates (Greiner, Kremsmünster, Austria) 24 h after siRNA transfection. To generate sphere formation, Spheromax CSC Medium (Promocell, Heidelberg, Germany) was added. After 7 days, sphere formation was assessed macroscopically. Additionally, sphere area in μm2 was quantified microscopically. Here, we photographically compared four microscopic fields for controls and knockdown cells. For area quantification, we used a previously published ImageJ Macro (Ivanov et al. 2014 (link)).
3
Sphere Formation Assay for CSCs
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Equal numbers of control pre-miR-and pre-miR-142-3p-transfected cells were seeded 24 h after transfection using coated six-well plates (Greiner, Kremsmu¨nster, Austria). Spheromax CSC Medium (4 mL) from Promocell (Heidelberg, Germany) was added. Spheres exceeding 50 mm in size were counted microscopically 8 days after seeding and normalized against the number of originally implanted cells. Spheroids of the first generation were trypsinized and seeded as single cells in a 96-well plate. Again, spheroids exceeding 50 mm in size were counted after 8 days of incubation.
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