DMEM/F-12 PLUS Basal Medium

3D Cell Culture Media BT-549 cells-Mammospheres

Experiment
3D Cell Culture Media BT-549 cells-Mammospheres
Product
DMEM/F-12 PLUS Basal Medium from Sigma-Aldrich
Manufacturer
Sigma-Aldrich

Protocol tips

Upstream tips
You can develop chemoresistant cell lines
Protocol tips
Seed 1.0 ×104 cells/well
Use of 24-ultralow attachment plates
Analysis of spheres with a diameter >40 μm
Downstream tips
Collect spheres for other procedures, such as protein analysis

Publication protocol

BT-549 cells (1.0 ×104 cells/well) were plated in 24-ultralow attachment plates (Corning Incorporate) and grown in DMEM/F-12 serum-free medium (Sigma-Aldrich), supplemented with B27 (1:50, Invitrogen), 20 ng/ml basic fibroblast growth factor (bFGF, Sigma- Aldrich) and 20 ng/ml EGF. After 4 days, the size and number of spheres were analyzed under a phase-contrast microscopy (Leica DMI3000 B apparatus). All spheres with a diameter >40 μm were analyzed, and the number and size of spheres were quantitated in at least 10 fields per condition. At the end of treatments, mammospheres were collected by gentle centrifugation (1000 rpm) and RNA was recovered using TRIzol reagent and then processed for RT-qPCR.

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Papers

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for DMEM/F-12 PLUS Basal Medium below.

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