Myelinating cocultures of rat retinal ganglion cell reaggregates and optic nerve oligodendrocyte precursor cells

Cold Spring Harb Protoc. 2014 Oct 1;2014(10):pdb.prot074971. doi: 10.1101/pdb.prot074971.

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.

MeSH terms

  • Animals
  • Antigens / metabolism
  • Axons / physiology
  • Cell Differentiation / drug effects
  • Cells, Cultured
  • Coculture Techniques*
  • Dendrites / physiology
  • Glial Fibrillary Acidic Protein / metabolism
  • Microtubule-Associated Proteins / metabolism
  • Myelin Basic Protein / metabolism
  • Myelin Sheath / physiology*
  • Oligodendroglia / physiology*
  • Optic Nerve / cytology*
  • Proteoglycans / metabolism
  • Rats
  • Retina / cytology*
  • Retinal Ganglion Cells / cytology
  • Retinal Ganglion Cells / physiology*
  • tau Proteins / metabolism

Substances

  • Antigens
  • Glial Fibrillary Acidic Protein
  • MAP2 protein, rat
  • Microtubule-Associated Proteins
  • Myelin Basic Protein
  • Proteoglycans
  • chondroitin sulfate proteoglycan 4
  • tau Proteins