Dulbecco’s Modified Eagle’s Medium/Ham’s Nutrient Mixture F12

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	Use of Ultra-Low Attachment 6-well plates

Protocol tips
Use of Ultra-Low Attachment 6-well plates

Publication protocol

Sphere formation assay: HLF and PLC/PRF/5 cells were trypsinized and cultured in DMEM-F12 (Sigma-Aldrich; Merck Millipore) supplemented with epidermal growth factor (Wako Pure Chemical Industries, Ltd., Osaka, Japan) at 20 ng/ml, basic fibroblast growth factor (Wako Pure Chemical Industries, Ltd.) at 20 ng/ml, 1% B27 supplement (Thermo Fisher Scientific, Inc.) and 1% methylcellulose 25 (Wako Pure Chemical Industries, Ltd.) in Ultra-Low Attachment 6-well plates (Corning Incorporated, Corning, NY, USA). The cells were imaged under a microscope (Olympus Corporation, Tokyo, Japan) after 7 and 14 days of incubation.

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Dulbecco’s Modified Eagle’s Medium/Ham’s Nutrient Mixture F12 below.

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