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Protocols: ES Cell Culture - Gene Targeting - Protocols

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