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(d) In neurosphere suspension culture condition, primary HPCa202N basal cells generated spheres more efficiently than luminal cells.
When the concentration was fixed at 0.1 M, the average diameter of the generated spheres is ~93 nm (see Fig. 1f), with a size distribution of standard deviation of ~11%.
Twelve of the 55 tumors yielded cells forming spheres that upon dissociation either did not generate novel spheres or generated spheres in numbers smaller than or similar to the ones evaluated in the previous passage.
A site box and the generated spheres were then prepared using the showbox and showsphere modules.
In contrast, 17 of 34 samples resistant to cisplatin generated spheres.
They were clonogenic and generated spheres in non-adhesive cell culture medium.
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A sphere ROI 6 mm in radius was built centered around each coordinate (using the MarsBaR toolbox for SPM); this size was chosen because it generated sphere ROIs generally comparable (or slightly smaller) in size with the clusters reported in these VBM studies, and this size of sphere ROI was commonly adopted in the literature employing similar analysis strategy.
If any newly generated sphere intersects with an existing sphere, the new sphere is rejected and another sphere with a different center and radius is generated.
CD44v3highALDH1high cells dissociated from spheres were counted by hemacytometer and replated to generate spheres of next generation.
In particular, the localization of binding site(s) is composed of three related operations: 1) generating the molecular surface of the receptor using the DMS program; 2) generating spheres surrounding the receptor using the Sphgen program; and 3) selecting a subset of spheres to represent the binding site(s) using the Sphere_selector program.
That is, an initial number of cells are seeded (e.g., 2.5×105), which generate spheres and produce a total cell count for the flask (e.g., 1×106), a portion of which are harvested (e.g., 25% are taken) and used to seed another flask for passage.
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