Dissociation, fractionation, and culture of embryonic brain cells

Abstract

Cell suspensions from 11-day chick embryo cerebral hemispheres were prepared by mechanical dissociation and cultured in a modified plasma clot system. Three distinct cell types are recognized: large cells (A), with the morphological characters of adult neurons, small cells (B) also presumably neuronal in character and thin, spread-out cells (C) which are epithelial. The neuronal-like elements, A and B, do not divide and have little mobility in culture: they grow very early, complexe processes which establish physical contact with one another and with other cell bodies. The epithelial cells, C, grow in culture even on uncoated glass surfaces, are very mobile, proliferate actively and can be propagated over a period of months. The A and C type cells have been fractionated from the harvested suspension by differential floation and selective glass attachment techniques to produce over 90% pure populations. The purification of the B type cells has not yet been achieved, due to their high and specific adhesiveness, both in suspension and in culture, to the C cells.