Figure 3.

Lineage commitment of hCNS-SCns derived cells in athymic nude (ATN) rat spinal cord injury (SCI) model at 14 weeks post-transplantation (WPT). It was found that 99.1% of human cells in the 9 DPI cohort were positive for either glial fibrillary acidic protein, doublecortin, or Olig2 at 14 WPT, whereas this combination of markers accounted for only 82.5% of human cells in the 60 DPI cohort (A). SC121/Olig2/APC (CC-1) triple immunostaining revealed that a significant fraction of human cells in the 60 DPI cohort were positive only for the mature oligodendroglial marker CC-1, and quantification of SC121⁺/CC-1⁺/Olig2⁻ cells in the 60 DPI transplant cohort raised the proportion of human cells exhibiting expression of lineage differentiation markers to be equivalent to the fraction detected in the 9 DPI hCNS-SCns cohort (A). (B–E): xyz projections show the 60 DPI cohort ATN rat spinal cord tissue at 14 WPT after immunohistochemistry using the human-specific cytoplasmic markers SC121 (B) and Olig2 (C) and the mature oligodendrocyte marker APC-1 (CC-1) (D) combined with nuclear counterstaining Hoechst. The overlay (E) demonstrates that some SC121⁺ human cells expressed both nuclear Olig2 and CC-1 (turquoise arrowhead), whereas others were positive for the mature oligodendrocyte marker CC-1 only (white arrowhead). Scale bar = 25 μm. Abbreviations: DPI, days postinjury; hCNS-SCns, human central nervous system-derived neural stem cell; N/A, not assessed.