PROLIFERATION AND DIFFERENTIATION OF FETAL HUMAN OLIGODENDROCYTES IN CULTURE

Phenotypic expression and proliferative capacity of the cells of oligodendrocyte lineage were investigated in primary cultures isolated from fetal human brains of 12-15 weeks' gestation using double immunolabeling with Ranscht-monoclonal antibody (R-mAb) or O4 and antibromodeoxyuridine (BrdU) antibody. Cultured cells of oligodendrocyte lineage consisted of a major population of R-mAb(+)O4(-) cells and minor populations of R-mAb(-)O4(+) and R-mAb(+)O4(+) cells. Most of the R-mAb(+)O4(-) cells exhibited a uni-, bi-, or tripolar immature morphology, while the majority of the R-mAb(+)O4(+) cells exhibited a multipolar mature morphology. R-mAb(-)O4(+) cells contained a mixture of immature and mature cell types. When incubated in serum-free culture medium containing BrdU for 4 days, 42% of total oligodendrocytes expressed nuclear BrdU immunolabeling. R-mAb(+) cells exhibited a higher degree of BrdU immunolabeling, indicating that they have greater capacities for proliferation than O4(+) cells. The large majority of BrdU(+) cells exhibited an immature morphology. Inclusion of insulin, insulin-like growth factor (IGF)-I, basic fibroblast growth factor (bFGF), or fetal bovine serum in culture medium did not stimulate proliferation of oligodendrocytes, while platelet-derived growth factor (PDGF) or PDGF plus bFGF increased the number of R-mAb(+)BrdU(+) and O4(+)BrdU(+) cells over control, even though the results were not statistically significant. In addition, insulin and IGF-I induced a 3-fold increase in the number of R-mAb(+)O4(+) cells, indicating that they promoted differentiation of oligodendrocytes. The present study indicates that fetal human oligodendrocytes in culture exhibit a considerable degree of proliferative capacity without requirement of exogenous growth factors and that both insulin and IGF-I promote their differentiation. (C) 1994 Wiley-Liss, Inc.