Oligodendrocytes are the myelinating cells of the central nervous system. Although the CNS possesses the ability to repair demyelinating insults, in certain cases, such as the chronic lesions found in multiple sclerosis, remyelination fails. Cycling cells capable of becoming oligodendrocytes have been identified in both the developing and the adult mammalian forebrain. Many studies have focused on differences in gene expression profiles as oligodendrocyte progenitors differentiate into myelinating oligodendrocytes by isolating cells at different developmental stages from animals at a single age. However, few have studied the differences that exist between the progenitors of the neonatal CNS and those of the adult CNS. This study examined the response of neonatal and adult O4(+) cells to platelet-derived growth factor-AA, basic fibroblast growth factor, and insulin-like growth factor-1 and revealed marked differences. Whereas adult cells readily differentiated in vitro, the majority of neonatal progenitors remained immature. Microarray analysis was used to examine differences between acutely isolated neonatal and adult progenitors further. Gene expression profiles showed that the adult O4(+) cells are more developmentally mature than neonatal cells. Neonatal cells expressed higher levels of genes involved in proliferation. Adult O4(+) cells expressed higher levels of transcripts for genes involved in cell death and survival. Therefore, O4(+) cells from the adult differ greatly from those of the neonate, and the developmental stage of the animal models utilized must be taken into consideration when applying principles from neonatal systems to the adult.