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Fig. 5. LIF maintains GFAP-expressing multipotent progenitors. (A) LIF and low (1-2
ng/ml) EGF propagate neural progenitor cells as a monolayer for at least 10
passages. (B) At higher passages (p7 is shown), GFAP-expressing cells
proliferate, as shown by BrdU incorporation (green, 2-hour pulse), and exhibit
the progenitor marker LeX (red). (C) Higher passage cultures are still able to
generate neurons (green), whereas nearby GFAP-expressing cells remain in cell
cycle, as demonstrated by Ki67 expression (red). (D,E) To directly test the
ability of GFAP-expressing cells for self-renewal and multipotentiality,
higher passage cells were infected with the rGFAPp-EGFP retrovirus, selected
by FACS, plated in a neurosphere-forming assay to assess self-renewal (in the
absence of high mitogen), and subsequent spheres plated for differentiation to
assess the ability to form neurons (red) and astrocytes (blue). (D)
GFAP-expressing cells are able to self-renew at high passages, as shown by
neurosphere formation (green, GFP). (E) Spheres formed from high passage
GFAP-expressing cells retain the ability to produce neurons (red; blue
indicates astrocytes; green, GFP). Scale bars: 20 µm in A-C,E; 100 µm in
D.
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