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. 2013 Jun 19;8(6):e67153. doi: 10.1371/journal.pone.0067153

Figure 2. Sphere formation and differentiation from the sciatic nerve in suspension culture.

Figure 2

(A) The sciatic nerve was cultured in DMEM-F12 medium containing bFGF. The ND-GFP-expressing cells proliferated and formed spheres by day 32. Bar: 100 µm. (B) The spheres expressing ND-GFP co-expressed p75NTR and CD34 but did not express β-III tubulin S100 and GFAP. Bar: 100 µm. (C) The ND-GFP-expressing spheres were switched to RPMI 1640 medium containing 10% FBS from DMEM-F12 containing B-27, N2 and bFGF and began to differentiate. At 7 days after switching into medium containing FBS, β-III tubulin-positive neuronal cells which expressed ND-GFP were observed. Bar: 10 µm. (D) At 14 days, the ND-GFP-expressing cells differentiated to GFAP-positive glial cells. Bar: 10 µm (E) At 7 days after culture in FBS medium, the ND-GFP-expressing cells differentiated to K15-positive keratinocytes, some of which still expressed nestin. Bar: 10 µm. (F) At 30 days after culture in FBS medium, the ND-GFP-expressing cells differentiated to α-SMA-positive smooth muscle cells. Bar: 50 µm.