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Components and Supplementation of Various Culture Media Abbreviations: DMEM = Dulbecco's Modified Eagle's Medium FCS = Fetal Calf (or Bovine) Serum GPS = Glutamine-Penicillin-Streptomycin BME = b-Mercaptoethanol DMSO = Dimethylsulfoxide HEPES = [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid] Buffer NEAA = Non-essential Amino Acids N = Nucleosides Feeder/STO Medium DMEM 7% FCS 1X GPS M15 (ES Cell Growth Medium) DMEM 15% FCS 1X GPS 1X BME 2X Freezing Medium This should be freshly prepared for each use. Wear gloves when working with DMSO. 60% DMEM 20% FCS 20% DMSO Injection Medium DMEM 5% FCS 20 mM HEPES pH 7.2-7.4 Embryo Culture Medium DMEM 20% FCS 1X GPS 1X BME 1X NEAA 1X N From the Laboratory of Dr. Allan Bradley Baylor College of Medicine, Houston, Texas