Figure 5. Immunostaining for hNSC reveals immature and mature cellular phenotypes independent of gender or hNSC labeling status.

A1) Immunostaining for human GFAP (hGFAP) identified positive (+) cells in the ipsilateral striatum (Str) adjacent to the lesion that had a mature astrocytic morphology distinct from those seen adjacent to the 3^rd ventricle. A2) hGFAP+ cells were also observed within the periventricular region, the median eminence (ME) and the arcuate nucleus (Arc), with fewer hGFAP+ cells were seen in the ventro-medial hypothalamus (VMH). hGFAP+ cells lining the 3^rd ventricle had similar morphology to astrocytic radial glia. B1) human-nestin (hNestin) within the contralateral cortex revealed hNSC that exhibited an intermediate filament morphology (arrows). B2) In the ipsilateral hemisphere, hNestin staining was observed adjacent to the ventricle (V), as well as tissues exposed to cerebrospinal fluid, such as cystic regions of the lesion (L), consistent with a more immature cellular phenotype, with only ~50% of these cells nestin+. C1–3) hNestin+ staining (arrows) along the ventricles did not colocalize with hGFAP+ cells (arrows). D1) Abundant endogenous rodent (rGFAP) astrogliosis (X) was observed in the ipsilateral cortex adjacent to the HII lesion. Higher magnification revealed numerous rGFAP+ astrocytes. D2) In contrast, only scattered hGFAP+ cells were observed within the cortical regions adjacent to the HII lesion. High magnification illustrated typical astrocyte morphology in an hGFAP+ cell. D3) No colocalization of human (green) and rat (red) astrocytes was observed. Higher magnification illustrates a single human astrocyte (green) surrounded by rodent astrocytes (red) with no colocalization. (cal bar – 20um)