Ectoderm and mesoderm interactions in the limb bud of the chick embryo studied by transfilter cultures: cartilage differentiation and ultrastructural observations.

The wing mesoderm of the chick embryo cultured in vitro without ectoderm is able to differentiate into cartilage from stage 17 (Hamburger & Hamilton, 1951). But before this stage the presence of ectoderm is necessary. In transfilter cultures of wing-bud ectoderm and mesoderm, the mesodermal response as measured by chondrogenesis was directly related to the pore size (0.2--1 micrometer) of the filter. Filters of 0.2 micrometer pore size and 10 micrometer thickness gave no increase in chondrogenesis over that of mesoderm cultures alone. The lower face of filters on the upper face of which mesoderm or ectoderm has been cultured was observed by scanning electron microscopy. With ectoderm, no cell processes crossed the filter. In contrast, with mesoderm, cell processes crossed the filter and this was also related to pore size. A good correlation was observed between the mass and density of processes crossing the filter and the mesodermal response. It is concluded that induction of cartilage in limb mesoderm cannot be classified as a 'long-range transmission' system. It requires ectoderm and mesoderm to be separated by a very narrow gap and this condition can be brought about in vitro by extension of mesodermal processes through the filter close to the ectoderm. The results are discussed in relation to a possible role of the basement membrane and associated extracellular matrix in limb cartilage induction.