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Figure 2. Spheroplasts. Treatment of E. coli with lysozyme which degrades the peptidoglycan layer in the presence of ethylene-diametetraacetate (EDTA) to permeabilize the outer membrane leads to the formation of osmotically fragile cells (spheroplasts) that are stable under hypertonic conditions. (i.e., 20-30% sucrose). Alternatively, cells can be grown in the presence of penicillin which inhibits peptidoglycan synthesis