The purpose of this work was to study the human fetal esophagus maintained in organ culture. Esophageal explants from 8 fetuses aged from 12 to 16 weeks of gestation were cultured up to 21 days at 37°C in Leibovitz L-15 serum-free medium. Between 12 and 16 weeks of gestation, the esophagus has a stratified columnar ciliated epithelium, and glycogen aggregates are present in all cell layers. This morphology remains the same up to 5 days in culture. After 7 to 9 days, a vacuolization in the upper half layer occurs, leading to a lifting off of the ciliated layer and a flattening of the subjacent cells. After 15 days of culture, the esophageal epithelium is stratified squamous and the cells are exfoliated at the surface of the explants. Glycogen aggregates are still present in all layers. Islets of ciliated cells resting on the basal cell layers develop within the squamous epithelium. With the extension of the culture period up to 21 days, the general morphology of the epithelium does not change. The ultrastructural features of the newly formed squamous epithelium, with its basal lamina, are similar to that reported for human adult esophageal epithelium. During the course of the culture, the DNA synthesis continues as determined by autoradiography. It is concluded that it is possible to maintain viable human fetal esophagus in organ culture and that an accelerated maturation takes place leading to the formation of the adult esophageal epithelium.