Neural stem cells from the central nervous system have the distinct capacity to give rise to clonal neurospheres. These clonal spheres are derived from a single clone-forming cell and represent homogenous, pure cell colonies. Recently, stem/progenitor cells have been isolated from the spiral ganglion of the inner ear using sphere-forming assays. However, the clonality of spiral ganglion-derived spheres has not yet been addressed in detail. Here, we report the isolation of clonal colonies from the spiral ganglion of early postnatal mice. We analyze sphere clonality using coculture experiments with transgenic cells, a semisolid assay, and culture of single cells in isolation. Our data show that sphere clonality differs in primary and secondary cultures and indicate that clonal sphere formation is dependent on specific culture parameters. We also show that the initiation of clonal colony formation does not require cell-to-cell interactions or paracrine signaling from surrounding cells. Generation of clonal colonies from spiral ganglion stem/progenitor cells might be crucial for future clinical applications because pure cell populations are considered to be more efficient and safe for therapeutic use than chimeric, heterogeneous spheres.