Figure 9. Single-cell clonal analysis of piglet OB- and forebrain-SVZ NSCs and genetic tagging with green fluorescent protein (GFP).

A. A single cell isolated from a neurosphere derived from the OB SVZ. Scale bar = 12 µm. B. Secondary neurosphere formed from neurosphere-derived single cells. Scale bar = 12 µm. C and D. FGF2/EGF-stimulated growth assays on dissociated neurosphere cells isolated from the forebrain-SVZ and OB-SVZ. The number of secondary neurospheres formed (C, values are mean ± SD) and individual neurosphere cell number (D) were assessed. Asterisk denotes SVZ-forebrain significant difference (p < 0.01) compared to SVZ-OB. E. Numerous piglet OB-SVZ-derived neurospheres were robustly and stably transduced to express GFP by lentiviral gene transfer. Inset shows a single GFP-tagged neurosphere at higher magnification. Some individual cells can be seen in the sphere. Scale bars: 80 µm, 5 µm (inset).