DMEM/F-12

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Publication protocol

Tumor-Sphere Culture. Hepatoma tumor spheres were generated and expanded in DMEM: F12 (Invitrogen) supplemented with B-27 (GIBCO) bFGF and EGF (R&D) in culture medium containing 1% methyl cellulose to prevent cell aggregation. Five hundred cells per well were seeded in 96 ultra-low attachment plates (Corning). The first generations of 10-day-old spheres were collected by gravity and dissociated to a single cell suspension with trypsin for serial passaging (G1-G5).

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Manufacturer protocol

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