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Fig. 4. In vitro differentiation of marrow-derived neurosphere-like structures into astroglial, oligodendroglial and neuronal cell types. hmNSCs were differentiated after 5-30 population doublings using the glial induction or the neuronal induction protocol on poly-L-lysine coated coverslips for 14 days. (A) hmNSCs differentiated using the neuronal induction medium were stained for markers for astrocytes (GFAP), oligodendrocytes (GalC), neurons (ß-tubulin-III, MAP2ab), or catecholaminergic cells (TH). Nuclei are counterstained with DAPI (blue). Bars, 50 µm. (B) Quantification of 14-day cultures of hmNSCs differentiated with the glial and the neuronal induction protocols. Data shown are mean±s.e.m. from at least three independent hMSC preparations. *, P<0.05; **, P<0.01 when compared to the percentage of positive cells in the glial induction protocol. (C) Clonal expansion from single hmNSCs. Six representative photomicrographs of neurosphere-like structures derived from single hmNSCs (left). Bar, 50 µm. Differentiation capacity of clonally derived neurosphere-like structures (right panel). Progeny of single cell-derived neurosphere-like structures can be differentiated into neurons (ß-tubulin III+, green) and astrocytes (GFAP+, red). Nuclei are counterstained with DAPI (blue). Bar, 30 µm.
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