GFP+ cells derived from a passaged neurosphere implant contribute to peripheral nerves and DRG. (A) After the removal of an ∼1-mm section of spinal cord,

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GFP+ cells derived from a passaged neurosphere implant contribute to peripheral nerves and DRG. (A) After the removal of an ∼1-mm section of spinal cord, the neurospheres were implanted in the lesion. GFP+ cells derived from a passaged neurosphere implant contribute to peripheral nerves and DRG. (A) After the removal of an ∼1-mm section of spinal cord, the neurospheres were implanted in the lesion. (B) After 1–2 wk of wound healing, the implanted neurospheres were integrated in the spinal cord tissue. The tail was amputated close to the integrated GFP+ cells, maintaining them within the 500-μm zone. (C) The integrated GFP+ cells contribute to spinal cord regeneration upon amputation. (D) MBP immunostaining of a tail cross-section containing regenerated EGFP+ spinal cord derived from the passaged neurosphere implant. An MBP+/EGFP+ nerve is shown in the box. (E) Confocal image of peripheral nerve showing GFP+ nuclei and MBP+/GFP+ myelin. (F) An anti-MBP–stained nerve myelin sheet. (G) GFP channel view of the nerve indicates EGFP+ cell nuclei and myelin cytoplasm. Green indicates EGFP+ cells, red indicates MBP immunostaining, and blue indicates Hoechst staining. (H) βIII-Tubulin immunostaining of the same cross-section. A DRG containing βIII-tubulin+/GFP+ cells is shown in the box. (I–K) Confocal image of DRG showing βIII-tubulin+/GFP+ cell and nerves. Green indicates EGFP+ cell bodies and nerves; red indicates βIII-tubulin immunostaining; blue indicates Hoechst staining. (Scale bars: D and H, 50 μm; E, F, and I–K, 10 μm.)‏ Levan Mchedlishvili et al. PNAS 2012;109:34:E2258-E2266 ©2012 by National Academy of Sciences