Characterization and development of glial and other non-neuronal cells in chick Edinger Westphal cultures

Abstract

Cultures of dissociated Edinger Westphal nuclei, dissected from embryonic chick brainstems, were screened immunohistochemically for a variety of non-neuronal cell markers. In young cultures, small clusters of cells were stained by the oligodendrocyte-specific antibodies 04 and 01. In older cultures, larger groups of cells were 04 and 01 positive, sheets of myelin-like membrane were elaborated, and immunoreactivity for proteolipid protein appeared. This sequence resembles that observed in well-characterized rodent brain cultures and suggests that oligodendrocytes in chick Edinger Westphal cultures differentiate in a pattern similar to rodent oligodendrocytes in culture. Variable numbers of cells were immunoreactive for glial fibrillary acidic protein. Many vimentin positive cells were observed, some of which morphologically resembled flat astrocytes. Together with the widespread presence of vimentin, large patches of fibronectin-like immunoreactivity suggested the presence of fibroblasts and/or endothelial cells. An anti-thymocyte polyclonal antibody stained a subset of cobblestone-shaped cells, possibly endothelial cells, in both Edinger Westphal cultures and control cultures of skin fibroblasts. Staining for smooth muscle myosia was detected in several patches of cells, tentatively identifying them as pericytes or smooth muscle cells. In conclusion, Edinger Westphal cultures contain a diverse and varying population of non-neuronal cells loosely organized in large, overlapping islands of cell types and including oligodendrocytes, astrocytes, possibly fibroblasts, endothelial cells, pericytes and/or smooth muscle cells.