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FLORA DE PABLO Y COLS. AN. R. ACAD. NAC. FARM.

FIGURE 5. Isolation of neural stem cells from E15 mouse olfactory bulb (O.B.).
Mouse embryo O.B. are dissected from brain (line over the dorsal view indicates the
section plane) and dissociated cells are placed in culture in the presence of FGF-2 and
EGF plus insulin or IGF-I. The cells divide actively and after 3-4 days they form
round aggregates, denominated neurospheres. After multiple passages the cells main-
tain their proliferative nature. When the mitogens are withdrawn (in the presence of
insulin or IGF-I) and cells are plated at high density, they enter a differentiation
program and, after 3-5 days, cells with the characteristics of neurons (TUJ1+), astro-
cytes (GFAP+) and oligodendrocytes (O4+) can be identified in the culture. The percen-

tages indicated are the average values obtained in a typical culture (40).

to E14.5 mice (40) (Fig. 5). These cells, growing as neurospheres,
were stimulated to proliferate by exogenous IGF-I in a dose-

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