Contralaterally transplanted human embryonic stem cell-derived neural precursor cells (ENStem-A) migrate and improve brain functions in stroke-damaged rats

Abstract

The transplantation of neural precursor cells (NPCs) is known to be a promising approach to ameliorating behavioral deficits after stroke in a rodent model of middle cerebral artery occlusion (MCAo). Previous studies have shown that transplanted NPCs migrate toward the infarct region, survive and differentiate into mature neurons to some extent. However, the spatiotemporal dynamics of NPC migration following transplantation into stroke animals have yet to be elucidated. In this study, we investigated the fates of human embryonic stem cell (hESC)-derived NPCs (ENStem-A) for 8 weeks following transplantation into the side contralateral to the infarct region using 7.0T animal magnetic resonance imaging (MRI). T2- and T2*-weighted MRI analyses indicated that the migrating cells were clearly detectable at the infarct boundary zone by 1 week, and the intensity of the MRI signals robustly increased within 4 weeks after transplantation. Afterwards, the signals were slightly increased or unchanged. At 8 weeks, we performed Prussian blue staining and immunohistochemical staining using human-specific markers, and found that high percentages of transplanted cells migrated to the infarct boundary. Most of these cells were CXCR4-positive. We also observed that the migrating cells expressed markers for various stages of neural differentiation, including Nestin, Tuj1, NeuN, TH, DARPP-32 and SV38, indicating that the transplanted cells may partially contribute to the reconstruction of the damaged neural tissues after stroke. Interestingly, we found that the extent of gliosis (glial fibrillary acidic protein-positive cells) and apoptosis (TUNEL-positive cells) were significantly decreased in the cell-transplanted group, suggesting that hESC-NPCs have a positive role in reducing glia scar formation and cell death after stroke. No tumors formed in our study. We also performed various behavioral tests, including rotarod, stepping and modified neurological severity score tests, and found that the transplanted animals exhibited significant improvements in sensorimotor functions during the 8 weeks after transplantation. Taken together, these results strongly suggest that hESC-NPCs have the capacity to migrate to the infarct region, form neural tissues efficiently and contribute to behavioral recovery in a rodent model of ischemic stroke.

Prolonged human neural stem cell maturation supports recovery in injured rodent CNS

“…This NSC line is well characterized: it expresses the NSC-specific markers nestin and SOX2 and differentiates into neurons, oligodendrocytes, and astrocytes (21). H9 NSCs were trans duced to express GFP and were harvested and embedded in fibrin matrices containing a growth factor cocktail (see Methods and ref.…”

Enhancing endogenous capacity to repair a stroke-damaged brain: An evolving field for stroke research

“…MSCs from cord blood, bone marrow and adipose decrease TUNELlabeled neurons, decrease caspase 3 mRNA, increase Bcl-2 expression (Calio et al, 2014; Feng et al, 2016; Li et al, 2002; Liu et al, 2014; Wu et al, 2008; Zhu et al, 2014). NSCs have similar effects when transplanted into the cortical peri-infarct area 24 h after permanent MCAO (Chen et al, 2014; Zhang et al, 2009) or even when administered one week after MCAO (Chang et al, 2013b; Shen et al, 2010. Even NSCs derived from iPSCs can decrease apoptosis (Chang et al, 2013a; Yao et al, 2015).…”

Transplantation of Adipose-Derived Stem Cells in Stroke

“…multi-lineage differentiation). Regarding the use of stems cells in the treatment of stroke, experimental studies using embryonic stem cells (Chang et al 2013;Drury-Stewart et al 2013), mesenchymal stem cells (Ikegame et al 2011), hematopoietic stem cells (Tsuji et al 2014), neural stem cells (Andres et al 2011), induced pluripotent stem cells (iPSC) (Oki et al 2012), and also multipotent adult progenitor cells (Mora- Lee et al 2012) in animal models of stroke have been reported with unanimous positive therapeutic results. Although mesenchymal stem cells were first identified four decades ago as adherent cells with fibroblastoid morphology being able to differentiate into cells of mesodermal origin such as osteocytes, chondrocytes, and adipocytes (Friedenstein et al 1974), they were later found to be also capable of differentiating into ectodermal and endodermal elements (Lakshmipathy and Verfaillie 2005;Ikegame et al 2011).…”