title: Clonal colony formation from spiral ganglion stem cells.
abstract: 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.
authors.affiliation: aDepartment of Otorhinolaryngology, Head and Neck Surgery, University Hospital Frankfurt am Main, Goethe University, Frankfurt/M., bInstitute of Veterinary Anatomy, Histology, and Embryology, Justus-Liebig University Giessen, Giessen, Germany.
authors.forename: Marc
authors.initials: M
authors.lastname: Diensthuber
authors.forename: Veronika
authors.initials: V
authors.lastname: Zecha
authors.forename: Jens
authors.initials: J
authors.lastname: Wagenblast
authors.forename: Stefan
authors.initials: S
authors.lastname: Arnhold
authors.forename: Timo
authors.initials: T
authors.lastname: Stöver
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002450:Cell Communication / Q000502:physiology; D018929:Cell Culture Techniques; D018920:Coculture Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D042282:Microdissection; D008856:Microscopy, Fluorescence; D058953:Neural Stem Cells / Q000502:physiology*; D013136:Spiral Ganglion / Q000502:physiology*; D000097573:Red Fluorescent Protein
chemical_list: D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein; C485184:enhanced green fluorescent protein