A NOVEL AND RAPID METHOD FOR CULTURING PURE RAT SPINAL-CORD ASTROCYTES ON UNTREATED GLASS

Astrocytes are the major population of glial cells, and an key players in the development, maintenance, and functioning of the central nervo us system (CNS). Their potential as targets of therapeutic interventio n following CNS injury makes the elucidation of their cellular and sub cellular physiology a primary research goal. Well defined and pure cel l culture systems are required to examine astrocytic physiology, bioch emical pathways and underlying responses to pathophysiologically alter ed conditions. Previously published protocols for establishing primary astrocyte cultures are time- and resource-consuming or suffer high co ntamination from other undesired cell types. Here we describe a new an d simple procedure for producing highly pure (> 99%) rat primary astro cyte cultures. The method involves a simple mechanical dissociation of harvested spinal cord tissue through a porous membrane and the subseq uent plating of the cells on plain, untreated glass coverslips. Astroc ytes adhere very well to the untreated glass while other cell types re quire a substrate such as poly-L-lysine. The method described here is, therefore, ideal for experiments which require highly pure astrocyte cultures. (C) 1998 Elsevier Science B.V. All rights reserved.