Mouse oligospheres: from pre-progenitors to functional oligodendrocytes.
To study the biology and repair capacities of mouse oligodendroglial cells, we established cultures of cells purified from neonatal wild-type and 9.6-kb MBP-LacZ transgenic newborn mice cerebral hemispheres as free-floating aggregates in the continuous presence of neuroblastoma conditioned medium (N1-B104). In vitro analysis indicated that the initial cell preparations were enriched in oligodendrocyte pre-progenitors that expressed PSA-NCAM and GAP-43 but not GD3, O4, NF68 or glial fibrillary acidic protein (GFAP) markers. These pre-progenitors required increased concentrations of insulin and progesterone to allow their survival in vitro. With time in culture, spheres composed of oligodendrocyte pre-progenitors became oligospheres enriched in oligodendrocyte progenitors expressing GAP-43 and GD3. As well as conserving bipotentiality in vitro, these spheres were able to form myelin in vivo after transplantation into the neonatal shiverer mouse brain. Thus, the oligosphere strategy is a powerful method for generating large populations of mouse oligodendrocyte pre-progenitors and progenitors. The ability to generate oligospheres from transgenic mice will be instrumental in the further dissection of the molecular and cellular mechanisms of myelination and remyelination of the central nervous system.
Vitry
S
S
Institut National de la Sante et de la Recherche Medicale CJF 97-11, Laboratoire des Pathologies de la Myeline, Paris cedex, France.
Avellana-Adalid
V
V
Hardy
R
R
Lachapelle
F
F
Baron-Van Evercooren
A
A
D000818:Animals; D002449:Cell Aggregation / Q000187:drug effects*; D018929:Cell Culture Techniques; D002490:Central Nervous System / Q000166:cytology / Q000196:embryology / Q000254:growth & development; D017077:Culture Media, Conditioned / Q000494:pharmacology; D019922:GAP-43 Protein / Q000032:analysis / Q000378:metabolism; D005699:Galactosylceramides / Q000032:analysis / Q000378:metabolism; D005732:Gangliosides / Q000032:analysis / Q000378:metabolism; D017930:Genes, Reporter; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis / Q000378:metabolism; D007004:Hypoglycemic Agents / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D007763:Lac Operon; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D039842:Neural Cell Adhesion Molecule L1; D019006:Neural Cell Adhesion Molecules / Q000032:analysis / Q000378:metabolism; D009836:Oligodendroglia / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D011374:Progesterone / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D012794:Sialic Acids / Q000032:analysis / Q000378:metabolism; D013234:Stem Cells / Q000187:drug effects* / Q000378:metabolism*
D017077:Culture Media, Conditioned; D019922:GAP-43 Protein; D005699:Galactosylceramides; D005732:Gangliosides; D005904:Glial Fibrillary Acidic Protein; D007004:Hypoglycemic Agents; D007328:Insulin; D039842:Neural Cell Adhesion Molecule L1; D019006:Neural Cell Adhesion Molecules; D012794:Sialic Acids; C002454:galactocerebroside; C103469:polysialyl neural cell adhesion molecule; D011374:Progesterone; C026226:ganglioside, GD3