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However, these methods are time consuming, cell spheres cannot be harvested easily, and it is difficult to control the size and geometry of cell spheres.
Formation of cell spheres is an important procedure in biomedical research.
A large number of high-quality cell spheres of uniform size and shape are required for basic studies and therapeutic applications.
To resolve these problems, a novel multiple-funnel cell culture insert was designed for size controlling, easy harvesting, and scale-up production of cell spheres.
To verify whether CART-HER2 cells can inhibit the growth of CSC subpopulations in primary GC, suspended cell spheres, which are aggregations of CSCs, were generated in serum-free media containing growth factors.
The formation of CD34/CD68/DLK cell spheres supported the interaction of ATMs, ASCs and preadipocytes.
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Conventional approaches, including the hanging drop method and suspension culture, are used for cell sphere production.
In contrast, when cells were cultured under stem cell (sphere) conditions, no disaggregation became apparent upon integrin inhibition, and cell death was not observed.
For the single cell sphere assay, single cells from the primary tumorspheres (1 cell/200 μl/well) were plated out in 96-well ultra-low attachment plates (Costar, Austin TX, USA) and spheres counted at 10 to 14 days.
Because each neurosphere was derived from a single cells, spheres were either entirely positive or negative for YFP (Figure 4A).
For experiments with adherent cells, spheres were dissociated and plated in N2 medium supplemented with 1% foetal bovine serum (FBS).
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