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American Journal of Pathology, Vol 83, 197-212, Copyright © 1976 by American Society for Investigative Pathology

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In vitro astrocytic differentiation from embryoid bodies of an experimental mouse testicular teratoma

SR VandenBerg, SK Ludwin, MM Herman and A Bignami

Astrocytic differentiation in monolayer cultures of ascitic embryoid bodies from the experimental teratoma OTT-6050 was studied by conventional light microscopy and by indirect immunofluorescence with antisera to glial fibrillary acidic (GFA) protein, a protein specific for astorcytes. Primitive neuroepithelial cells were identified in 24- hour cultures. Within 72 hours, two cell types diverged. One cell type, with a flattened epithelial morphology in early cultures, demonstrated delicate GFA protein-positive fibrils within 48 hours. In later cultures, this type progressively displayed more typical stellate astrocytic features, with denser, more compact GFA protein-positive fluorescence in the perinuclear cytoplasm and cell processes. As indicated by GFA protein expression, the appearance of astrocytes of typical morphology therefore was preceded by biochemical differentiation. The second cell type, interpreted as neuroblastic, failed to demonstrate GFA protein and had a small perikaryon with slender bipolar processes that were argyrophilic with Bodian's protargol in late cultures. Divergent neuroepithelial differentiation occurred within mitotically active cell populations and proceeded without apparent tissue relationships to other germ layer derivatives.