Clustering of central nervous system (CNS) cells is often utilized for cell growth and characterization, as well as investigated for tissue regeneration and disease progression. Collective CNS cell migration, however, has been largely unstudied. Cell cluster formation and migration play a critical part of modeling in vivo conditions and in development of therapies. Three distinct CNS cell types, medulloblastoma (MB), medulloblastoma-derived glial progenitor cells (MGPC), and retinal progenitor cells (RPC), were investigated for cluster formation, upregulation of CXCR4, the receptor for Stromal-Derived Growth Factor (SDF-1), and Connexin 43 expression, a gap junction hemichannel. A microfluidic platform was used to examine the the migration of clusters and single cells in response to controlled concentration gradients of SDF-1. All cell types illustrated self-clustering, as well as upregulated CXCR4 surface expression and increased Connexin 43 expression upon ligand stimulation. Further, RPC clusters exhibited collective, chemotactic migration along SDF-1 concentration gradients, while MB clusters illustrated inconsistent collective migration, and MGPCs clusters did not exhibit adhesion-based migration.