title: Myelinating cocultures of rat retinal ganglion cell reaggregates and optic nerve oligodendrocyte precursor cells.
abstract: This protocol describes the generation of a rapidly myelinating central nervous system coculture for the study of complex neuronal-glial interactions in vitro. Postnatal rat retinal ganglion cells (RGCs) purified by immunopanning are promoted to cluster into reaggregates and then allowed to extend dense beds of radial axons for 10-14 d. Subsequently, rodent oligodendrocyte precursor cells are purified by immunopanning, transfected if desired, and seeded on top of the RGC reaggregates. Under the conditions described here, compact myelin can be observed within 6 d.
authors.affiliation: Department of Neurobiology, Stanford University School of Medicine, Stanford, California 94305-5125.
authors.forename: Trent A
authors.initials: TA
authors.lastname: Watkins
authors.affiliation: Department of Neurobiology, Stanford University School of Medicine, Stanford, California 94305-5125.
authors.forename: Anja R
authors.initials: AR
authors.lastname: Scholze
mesh_terms: D000818:Animals; D000941:Antigens / Q000378:metabolism; D001369:Axons / Q000502:physiology; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D003712:Dendrites / Q000502:physiology; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D004676:Myelin Basic Protein / Q000378:metabolism; D009186:Myelin Sheath / Q000502:physiology*; D009836:Oligodendroglia / Q000502:physiology*; D009900:Optic Nerve / Q000166:cytology*; D011509:Proteoglycans / Q000378:metabolism; D051381:Rats; D012160:Retina / Q000166:cytology*; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000502:physiology*; D016875:tau Proteins / Q000378:metabolism; D000099216:Chondroitin Sulfate Proteoglycan 4
chemical_list: D000941:Antigens; D005904:Glial Fibrillary Acidic Protein; D008869:Microtubule-Associated Proteins; D004676:Myelin Basic Protein; D011509:Proteoglycans; D016875:tau Proteins; D000099216:Chondroitin Sulfate Proteoglycan 4; C489787:MAP2 protein, rat