MEGMTM Mammary Epithelial Cell Growth Medium BulletKitTM

Protocol tips

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	Seed BT549 cells at a density of 2500 cells/wel/ Use of 6-well ultra-low attachment plates Count mammospheres with diameters of ≥100 μm

Protocol tips
Seed BT549 cells at a density of 2500 cells/wel/ Use of 6-well ultra-low attachment plates Count mammospheres with diameters of ≥100 μm

Publication protocol

BT549 and MDA-MB-231 cells, when seeded in ultra-low attachment plates (Corning, Corning, NY) in serum-free media form non-adherent spheroids termed mammospheres, with the ability to self-renew (Fillmore and Kuperwasser, 2008). To examine the effects of various treatments on mammosphere formation, BT549 and MDA-MB-231 cells were seeded in 6-well ultra-low attachment plates in plating medium (MEGM) with and without added treatments at a density of 2500 cells per well. 24h after, chemotherapeutic agents were added into plate. The appearance of spheres was evaluated after 7 (BT-549) or 10-12 (MDA-MB-231) days. Mammospheres with diameters of ≥100 μm (BT-549) and ≥50 μm (MDA-MB-231) were manually counted under microscope.

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Manufacturer protocol

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