Microglia, astrocytes, and neurons, which have important functions in the central nervous system, communicate mutually to generate a signal through secreted proteins or small molecules, but many of which have not been identified. Because establishing a reference for the secreted proteins from CNS cells could be invaluable in examining cell-to-cell communication in the brain, we analyzed the secretome of 3 murine CNS cell lines without pre-fractionation by high-resolution mass spectrometry. In this study, 2795 proteins were identified from conditioned media of the 3 cell lines, and 2125 proteins were annotated as secreted proteins by bioinformatics analysis. Further, approximately 500 secreted proteins were quantifiable as differentially expressed proteins by label-free quantitation. As a result, our secretome references are useful datasets for the future study of neuronal diseases.