SCANNING ELECTRON-MICROSCOPIC OBSERVATIONS OF HUMAN FETAL KIDNEY MATURING INVIVO AND IN SERUM-FREE ORGAN-CULTURE

A serum-free model has been developed in our laboratory enabling us to maintain human fetal kidney in culture for periods of 5 days or more. In this totally defined system, morphological integrity of these explants was shown to be preserved at both the light and the electron microscopic levels. The present work was undertaken to validate our culture model via scanning electron microscopy, a technique allowing surface observation of micromorphological features overlooked by conventional microscopy. In uncultured kidney, different developmental stages of nephron formation were identified. A sparse population of short microvilli was present on most cell apical membranes. Cell outlines were polygonal and demarcated by longer and densely packed microvilli. In proximal tubules, these microvilli were in the process of forming a brush border. In the majority of cells, one or two cilia with twisted or hooked tips projected into the capsular space or tubule lumen. Microcraters and bleb-like structures characterized the luminal membrane of many cells. The urinary papilla epithelium was composed of some ciliated principal cells but mostly of intercalated cells with either apical microplicae, microvilli, or both. Microprojections formed zipper-like intercellular junctions. In culture, ultrastructural features, including membrane pits and spherical vesicles, were similar to those in uncultured explants. In summary, these novel observations in cultured fetal kidney indicate that ultrastructural integrity is well preserved in serum-free medium and that the present model is a valuable tool to study human nephrogenesis.