Glial cells derived from aged mouse brain in culture display both mature and immature astrocytic phenotypes

In earlier studies, we established glial cell cultures derived from aged (18-month-old) mouse cerebral hemispheres (MACH) and have maintained them frozen at various passages. These cultures were characterized immunocytochemically and consist of: 5% oligodendrocytes (GalC+), 75% astrocytes-type 1 (GFAP+ only), 15% astrocytes-type 2 (GFAP+ + A2B5+), and 5% progenitor glial cells (A2B5+ only). In the present study, we isolated colonies from MACH passage 29 cultures and also colonies from MACH passage 19 transfected with the gene for SV40 large T antigen and further subcultured for 8 passages. Using double-staining immunocytochemistry, we found in non-transfected MACH passage 19 colonies consisting primarily of cells exhibiting only vimentin-positive staining and are considered to be immature glioblasts; colonies consisting primarily of cells exhibiting GFAP+ + vimentin+ which are considered to be astrocytes at an intermediate stage of maturation; and colonies consisting predominantly of cells exhibiting GFAP+ only which are considered to be mature astrocytes. In contrast, colonies isolated from transfected MACH cultures consisted primarily of vimentin+ cells. In conclusion, astrocytes in cultures derived from aged brain continue to be variable as they are during development. However, their response to the microenvironment may differ during development and during aging. Thus, the availability of clones of mature and immature astrocytes offers the opportunity to study neuron-glia interactions and the role of mature and immature astrocytes in neuronal aging and regeneration.