MammoCult™ Human Medium Kit

Protocol tips

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	Seed at a density of 5,000 cells/mL Use 6-well ultra-low-attachment plates Count mammospheres ≥50 μm in diameter

Protocol tips
Seed at a density of 5,000 cells/mL Use 6-well ultra-low-attachment plates Count mammospheres ≥50 μm in diameter

Publication protocol

Cultured cells were trypsinized, whereas tumor tissues were minced, digested with 1 mg/mL of type 1 collagenase (Sigma-Aldrich) at 37°C for 30 min, and filtered through a 70-μm cell strainer. The number of live cells was determined using trypan blue staining. Single-cell suspensions were seeded in 6-well ultra-low-attachment plates (Corning) at a density of 5,000 cells/mL in complete MammoCult Medium (STEMCELL Technologies). Mammosphere cultures were photographed 7 days later using a phase contrast microscope (Olympus), and mammospheres ≥50 μm in diameter were counted using ImageJ software (NIH). Primary mammospheres were collected by centrifugation and dissociated using trypsin, and single-cell suspensions were re-seeded at a density of 2,500 cells/mL for the secondary mammosphere assay.

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Manufacturer protocol

Download the product protocol from STEMCELL technologies for MammoCult™ Human Medium Kit below.

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