RAPID PURIFICATION OF GLIAL-CELLS USING IMMUNOMAGNETIC SEPARATION

By purifying glial cells from brain tissue containing a heterogeneous cell population, a number of interactions that define glial cell diver sification and function within the central nervous system have been de termined. The current methods for purifying glial cells, however, can be time consuming and costly. In the following study we have adapted t he technique of immunomagnetic separation to separately enrich 0-2A pr ogenitor cells and astrocytes from the rat central nervous system (CNS ). In this procedure, tissue from the CNS was enzymatically dissociate d and incubated in a primary antibody specific to a surface antigen fo und on the target cell type (e.g. A2B5 or RAN-2). The target cells wer e then immunologically coupled to magnetic beads, which were precoated with a secondary antibody specific to the primary, and then separated out from the heterogeneous cell population using a magnetic field. We found that the immunomagnetic separation procedure, which was complet ed within 2 h, produced a near pure population of glial cells (>99%). This was confirmed by the absence of unbound cells in the bead-bound f raction. The identification and viability of bead-bound cells were est ablished by culturing these cells and subsequently examining their mor phology and antigenic expression. This study shows that glial cell typ es can be separated out of brain tissue to near purity using immunomag netic separation. This simple procedure is reliable, inexpensive, and achieves levels of purity and viability comparable with currently avai lable techniques of immunopanning and fluorescence-activated cell sort ing, within a fraction of the time. (C) 1997 Elsevier Science B.V.