Studies of glial lineage and proliferation in vitro using an early marker for committed oligodendrocytes

The potential of immature glial cells to differentiate into astrocytes (ASs) or oligodendrocytes (OLs) has been examined using a monoclonal antibody (007) that is specific for OLs in vivo. Cells were dissociated from 2-day postnatal mouse cortex and labeled with the 007 antibody 2 hr after plating. The cells which were labeled during this single, brief exposure to the antibody retained the antibody on their surfaces over the course of the experiments. Cells were double stained at various timepoints for residual 007 antibody and either galactocerebroside (GC) or glial fibrillary acidic protein (GFAP). Shortly after plating, most 007+ cells were GC- and none expressed GFAP. These cells were round, although some had begun to extend very short processes. After 96 hr, greater than 95% of cells with residual 007 on their surfaces also expressed GC. By this time, all the 007+ cells had several processes of varying lengths extending from their cell bodies. Cells expressing both 007 and GFAP were never seen. The 007+/GC+ OLs were not induced to differentiate from 007+ bipotential progenitors since they were grown in fetal calf serum. These results show that under our culture conditions the 007 antibody is OL specific. Immunostaining for bromodeoxyuridine, a marker for dividing cells, revealed that some 007+ cells were proliferating. The majority of these proliferating cells had already extended three or more processes. We therefore conclude that immature, process-bearing cells can be committed to the OL lineage at times before they express detectable amounts of GC. Since these young 007+ OLs are actively proliferating, committed cells can serve as an important source of new OLs.