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This protocol describes the feeder-dependent culture of hESCs. . MEFs on an Attachment Factor-coated culture vessel in MEF medium (refer to Table .. It is common practice to condition new feeder plates before passaging hESCs into them.
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Mouse Embryonic Stem Cell (mESC) Culture Medium (100 mL of complete medium) .. It is common practice to condition new feeder plates before passaging.
At what density should I plate my MEF feeder cells? . layer, the medium can be switched to stem cell medium when the stem cells are plated on the feeders.
MEF medium (see recipe) with penicillin/streptomycin. Laminar flow hood . using approximately one embryo per plate or flask. Add medium to a . MEFs must be inactivated prior to use as a feeder layer for mouse ES cells. Mitotic inactivation.
Remove DMSO containing freezing medium and resuspend in feeder medium. Plate appropriate number of MEF /size of plate needed on gelatinized plates.
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