Acceleration of the maturation of oligodendroblasts into oligodendrocytes and enhancement of their myelinogenic properties by a chemically defined medium.
Because of the importance of oligodendrocytes (OL) in forming and maintaining myelin in the CNS and the fact that the remyelination in the CNS is very limited in contrast to the peripheral nervous system, we investigated the effect of a chemically defined medium OLDEM, previously characterized by the maintenance of mature myelinating OL, on oligodendroblasts (or OL progenitors) in culture. The effect of each component of this medium as well as different combinations of them were also examined. Cultures were examined at different developmental stages immunocytochemically for developmental markers, such as transferrin, sulfatides, myelin basic protein and proteolipid protein. OLDEM accelerated the appearance of developmental markers and concomitant morphological changes. Furthermore, myelin-specific enzymes such as glycerophosphorylcholine phosphodiesterase; p-nitro-phenolphosphocholine phosphodiesterase; 2'3'-cyclic nucleotide 3'-phosphodiesterase and UDP galactose: ceramide galactosyltransferase had enzymatic activities similar to values found in pure myelin, indicating that OLDEM allows the optimal expression of myelin-related genes. The effect of each OLDEM constituent was evaluated by immunocytochemistry and by measurement of enzymatic activities. With each single additive or multiple combinations, oligodendrocytes displayed different degrees of maturation. Deletion of selenium, glucose, and galactose severely affected cell survival as well as enzymes expression in young cultures. However, older cultures were more resistant to these deletions. Putrescine and insulin did not cause such effects on survival, but their absence affected cell maturation. None of the OLDEM additives individually supported survival and/or maturation. Enzyme assays performed on isolated myelin-like membranes or the cells soma revealed a redistribution of the activity between these fractions as the cell matured. The biological role of each of these constituents on the maturation of the oligodendroglial cells is discussed. These observations indicate that OLDEM constituents have a powerful effect on OL progenitor maturation, and membrane formation. This medium will be used for investigating the remyelination potential of adult OL progenitors.
Espinosa de los Monteros
A
A
Department of Neurobiology, UCLA School of Medicine, USA.
Yuan
J
J
McCartney
D
D
Madrid
R
R
Cole
R
R
Kanfer
J N
JN
deVellis
J
J
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D003470:Culture Media; D007150:Immunohistochemistry; D009186:Myelin Sheath / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D009457:Neuroglia / Q000502:physiology; D009836:Oligodendroglia / Q000201:enzymology / Q000502:physiology* / Q000648:ultrastructure; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D051381:Rats
D003470:Culture Media; D010727:Phosphoric Diester Hydrolases