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SUMMARY
Protoplasts of the yeast phase of Histoplasma capsulatum prepared by incubation in Sabouraud broth + 1.5 M MgSO4 + 2-deoxy-d-glucose will regenerate to the sporulating mycelial phase in a variety of osmotically supportive media at 25 C, and to the budding yeast phase at 37 C. Freshly formed and isolated protoplasts are osmotically fragile. They become resistant to osmotic shock within a few hours at 4 C and remain viable and capable of regeneration after more than 2 months at -70 C. Methods of protoplast induction, separation and concentration under sterile conditions are described.