A simultaneous ex vivo model of embryogenesis: I. Organogenesis.

ABSTRACT

In vivo culture of chick embryo was carried out to develop an experimental interphase between in vitro and in vivo study of embryonic physiology. In the process, a simultaneous model of vasculogenesis and organogenesis has been worked out, which is impossible to achieve in mammalian system. Both early (40 hours of incubation) and late (64 & 88 hours of incubation) hours of cultures were conducted for morphological and morphometric studies. A new combination of stains was used in place of conventional haematoxylin and eosin in 40 hours old whole-mount of embryos. Semithin plastic sections were etched for haematoxylin/pyronin stain in addition to paraffin (both normal and enblock) sections. Specific stains (histological, enzyme histochemical or immunohistochemical) were chosen according to the specific organs/areas studied. Immunohistochemistry and NADPH-diaphorase activity were standardized in whole-mount of embryos. Morphometry was done using camera lucida and quantitative image analysis system. A parallel preparation of extra embryonic whole-mounts, paraffin and semithin plastic sections with different types of stainings provides evidence for the scope of the simultaneous study of vasculogenesis. Thus the morphological and morphometric data presented in this and the succeeding article describe the scope and avenues for the use of ex vivo model in various aspects of embryonic physiology, preliminary drug trials/metabolism, radiology, teratology and toxicology.