Concurrent isolation and characterization of oligodendrocytes, microglia and astrocytes from adult human spinal cord.
A cellular preparation of highly enriched oligodendrocytes was obtained from adult human spinal cord by Percoll gradient centrifugation followed by either differential adhesion or fluorescence-activated cell sorting after immunostaining with an antibody against galactocerebroside (O1). The adherent and O1-negative cell fractions were > 96% microglia. The non-adherent and O1-positive fractions were > 96% positive for the oligodendrocyte markers O4 and O1, 0-2% positive for glial fibrillary acidic protein, and were devoid of neuronal or microglial markers. If the oligodendrocyte fraction was co-cultured with purified dissociated rat dorsal root ganglion neurons, the oligodendrocytes adhered to the axons and their numbers increased over a 4 week period. However, myelin sheaths were not produced around axons in these cultures. In contrast, if the oligodendrocyte cell fraction was grown alone in culture for > 3 weeks, the number of oligodendrocytes decreased and a layer of astrocytes developed underneath the oligodendrocytes. The oligodendrocytes could be eliminated from these cultures by subsequent passaging, thus producing cultures of pure astrocytes. The astrocytes accumulated both K+ and glutamate with kinetic properties similar to those reported for rodent astrocytes. We suggest that these astrocytes arose in part from an O4/O1-positive precursor which did not initially express glial fibrillary acidic protein. These results define a relatively simple method by which highly enriched populations of oligodendrocytes, astrocytes and microglia can be obtained from adult human spinal cord.
Whittemore
S R
SR
Miami Project, University of Miami School of Medicine, FL 33136.
Sanon
H R
HR
Wood
P M
PM
D000293:Adolescent; D000328:Adult; D000906:Antibodies / Q000032:analysis; D001253:Astrocytes / Q000276:immunology / Q000502:physiology*; D002448:Cell Adhesion; D003584:Cytological Techniques; D005699:Galactosylceramides / Q000276:immunology / Q000378:metabolism; D005727:Ganglia, Spinal / Q000166:cytology; D005904:Glial Fibrillary Acidic Protein / Q000276:immunology / Q000378:metabolism; D005971:Glutamates / Q000378:metabolism; D018698:Glutamic Acid; D006801:Humans; D007150:Immunohistochemistry; D017628:Microglia / Q000276:immunology / Q000502:physiology*; D008875:Middle Aged; D009836:Oligodendroglia / Q000276:immunology / Q000502:physiology*; D011188:Potassium / Q000378:metabolism; D013116:Spinal Cord / Q000166:cytology*
D000906:Antibodies; D005699:Galactosylceramides; D005904:Glial Fibrillary Acidic Protein; D005971:Glutamates; D018698:Glutamic Acid; D011188:Potassium