Technical Procedures for the Study of Organogenesis in vitro in Amblystoma

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Abstract
Technical procedures have been developed which permit the study of organogenesis in vitro in explants from embryos of A. punctatum. An embryo from which the explant was to be taken was rendered sterile by placing it for 24 hrs. in a soln. of 0.5% or 1% Na sulfadiazine, plus 10-20 gamma/ml. of streptomycin. Using this procedure sterilization of embryos was accomplished in 95% of cases. The nutrient medium in which the explant grew consisted of 2 fluids: 1) citrated rabbit plasma, and 2) half-strength Holtfreter soln. to which was added 0.25% agar, 0.045% Na nucleate, and 0.77% glucose. Equal portions of the 2 fluids were mixed to make the final medium. Organogenesis occurred only when the explant became entirely covered by epidermis. Epibolic activity of the epidermis was fostered by immersion of the explant in full-strength Holtfreter soln. for 2 hrs. immediately before mounting in a hanging drop. Explants were kept at 20 C.
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