The production of extracellular connective tissue fibrils by chick notochordal epithelium in vitro

Abstract

Abstract Fertile hens eggs were incubated 48 hours. Embryos were removed from the shell and the cervical region from each excised and placed in a balanced salt solution containing 1% trypsin for 40 minutes. Notochords were isolated by microdissection and further incubated in vitro 48 or 72 hours. Following trypsinization, unincubated notochords were rod‐shaped and were shown by electron microscopy to be devoid of extracellular materials or contaminating mesenchymal cells. Extracellular materials, ultrastructurally indistinguishable from those in the perinotochordal connective tissue space in vivo, are present on the surfaces of cell clusters at 48 hours of in vitro incubation. Areas of fibrillogenesis, in which microfibrils are separated from the surface of cells by intermittent basement lamina, are common. At 72 hours of in vitro incubation fibrogenic activity is less evident, but occasionally intense concentrations of small and large microfibrils, basement lamina and other extracellular substances are seen adjacent to notochordal cell surfaces. These observations are of special interest in light of the known role of the notochord in embryonic induction and recent demonstrations that surface‐associated substances (specifically collagen) are necessary for normal cytodifferentiation.