Generation of Mouse Astroglia and Microglia Cultures from Mouse Neopallium

Abstract

Newborn rats or mice are generally used as the source of tissue for glial cultures. Only about 1% of cells survive the cell disaggregation process and culture environment, and neurons that survive die within the first few days of culturing. Terminally differentiated cells, which can no longer divide, are overgrown by the proliferating, immature cells. As a result, the cells in culture are mainly immature cells and therefore cultures are very plastic. How the cultures will develop and which cell types, i.e., astroglia, Oligodendroglia, ependymal cells, or microglia, will enrich the culture, depends on the culture medium and the physical conditions under which the cells are grown. In addition to varying the components of a chemically defined medium or adding or removing serum from the medium, it is possible to add to the cultures growth factors and/or cytokines in pure recombinant form, or as soluble products in medium conditioned by cells, that produce and secrete the factors. (The latter is considerably cheaper). The addition of such factors to cultures can have dramatic effects. It should be noted that such factors may affect more than one cell type and may initiate different effects in different cells. The factors may interact with other factors synergistically, additively, or in an inhibitory way. Moreover, their half-life is short. This subject is extremely complex and beyond the scope of this book.