Adherent Self-Renewable Human Embryonic Stem Cell-Derived Neural Stem Cell Line: Functional Engraftment in Experimental Stroke Model

Department of Neurosurgery and Stanford Stroke Center, Stanford University School of Medicine, Stanford, California, USA.
PLoS ONE (Impact Factor: 3.23). 02/2008; 3(2):e1644. DOI: 10.1371/journal.pone.0001644
Source: PubMed


Human embryonic stem cells (hESCs) offer a virtually unlimited source of neural cells for structural repair in neurological disorders, such as stroke. Neural cells can be derived from hESCs either by direct enrichment, or by isolating specific growth factor-responsive and expandable populations of human neural stem cells (hNSCs). Studies have indicated that the direct enrichment method generates a heterogeneous population of cells that may contain residual undifferentiated stem cells that could lead to tumor formation in vivo.
We isolated an expandable and homogenous population of hNSCs (named SD56) from hESCs using a defined media supplemented with epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and leukemia inhibitory growth factor (LIF). These hNSCs grew as an adherent monolayer culture. They were fully neuralized and uniformly expressed molecular features of NSCs, including nestin, vimentin and radial glial markers. These hNSCs did not express the pluripotency markers Oct4 or Nanog, nor did they express markers for the mesoderm or endoderm lineages. The self-renewal property of the hNSCs was characterized by a predominant symmetrical mode of cell division. The SD56 hNSCs differentiated into neurons, astrocytes and oligodendrocytes throughout multiple passages in vitro, as well as after transplantation. Together, these criteria confirm the definitive NSC identity of the SD56 cell line. Importantly, they exhibited no chromosome abnormalities and did not form tumors after implantation into rat ischemic brains and into naïve nude rat brains and flanks. Furthermore, hNSCs isolated under these conditions migrated toward the ischemia-injured adult brain parenchyma and improved the independent use of the stroke-impaired forelimb two months post-transplantation.
The SD56 human neural stem cells derived under the reported conditions are stable, do not form tumors in vivo and enable functional recovery after stroke. These properties indicate that this hNSC line may offer a renewable, homogenous source of neural cells that will be valuable for basic and translational research.

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    • "However, their identity as true radial glial cells that recapitulate their in vivo counterparts is debatable. Notwithstanding the phenotypic consistency of SENAs, an isolated human neural stem cell line termed SD56, expressing vimentin, nestin and 3CB2 (markers of early appearing radial glia (Prada et al., 1995)) showed extensive migration without tumorigenesis around a striatal ischaemic lesion in the rat, significantly improving the independent use of the stroke impaired forelimb (Daadi et al., 2008). Perhaps a more accurate paradigm of radial glial cell transplantation is the C6 glioma derived radial glia-like cell line C6- R. C6-R cells show a bipolar morphology in vitro and support neuronal migration, while expressing markers typical of in vivo radial glia including vimentin, nestin, glial fibrillary acidic protein (GFAP) and RC1 (Friedlander et al., 1998). "

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    • "Culturing condition might reduce tumorigenesis risk of transplanted ESC-derived neural cells. For example, neural cells derived from human ESCs under defined inductive culturing condition (named SD56) did not show chromosome abnormalities after differentiation and tumor formation after implantation into ischemic rat brains and naive nude rat brains and flanks [19]. Malignant transformation of ESC-derived neural cells has been demonstrated to be related to postischemic environment probably by the stimulation of various local cytokine [26]. "
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    ABSTRACT: In recent years, stem cell-based approaches have attracted more attention from scientists and clinicians due to their possible therapeutical effect on stroke. Animal studies have demonstrated that the beneficial effects of stem cells including embryonic stem cells (ESCs), inducible pluripotent stem cells (iPSCs), neural stem cells (NSCs), and mesenchymal stem cell (MSCs) might be due to cell replacement, neuroprotection, endogenous neurogenesis, angiogenesis, and modulation on inflammation and immune response. Although several clinical studies have shown the high efficiency and safety of stem cell in stroke management, mainly MSCs, some issues regarding to cell homing, survival, tracking, safety, and optimal cell transplantation protocol, such as cell dose and time window, should be addressed. Undoubtably, stem cell-based gene therapy represents a novel potential therapeutic strategy for stroke in future.
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    • "ESC studies in animal stroke models have been concerned with mechanistic aspects rather than functional efficacy, and report only isolation, neutralization,89 and the electrophysiological activity of differentiated neuronal cells.90 Undifferentiated ESCs grafted into rat brains have differentiated and integrated with host tissues in stroke models,91 showing improved functional outcomes on the cylinder test, which measures the spontaneous use of forelimbs.92 "
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