Experimental studies on the differentiation of embryonic tissues growing in vivo and in vitro .—I. The development of the undifferentiated limb-bud (a) when subcutaneously grafted into the post-embryonic chick and (b) when cultivated in vitro

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Abstract
Undifferentiated limb buds of fowl embryos of 82 hrs.'' incubation when inoculated subcutaneously into the under surface of the wings of 11-day post-embryonic chicks differentiated into cartilage, bone, fibrous tissue, and typical epidermis. Differentiated muscle was absent, and the general morphology of the skeletal tissue in the grafts bore apparently no relation to that of the normal limb skeleton. When cultivated in vitro by an improved tube technique the undifferentiated limb buds from embryos of 72-80 hrs.'' incubation showed active cell division and differentiation into cartilage, white fibrous tissue, and typical epidermis. When grown in a relatively fluid medium many of the cultures developed into large balloon-like cysts; in such cases the shape and arrangement of the cartilaginous nodules showed no correspondence to the structure of the developing cartilage bones of the normal limb. With a more solid culture medium, cyst formation was far less pronounced, and skeletogenous tissue frequently showed the early stages in the development of a normal limb bud.

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