ABSTRACT
Neural stem cells (NSCs) are multipotent, self-renewable cells who are capable of differentiating into neurons, astrocytes, and oligodendrocytes. NSCs reside at the subventricular zone (SVZ) of the adult brain permanently to guarantee a lifelong neurogenesis during neural network plasticity or undesirable injuries. Although the specious inaccessibility of adult NSCs niche hampers their in vivo identification, researchers have been seeking ways to optimize adult NSCs isolation, expansion, and differentiation, in vitro. NSCs were isolated from rhesus monkey SVZ, expanded in vitro and then characterized for NSCs-specific markers expression by immunostaining, real-time PCR, flow cytometry, and cell differentiation assessments. Moreover, cell survival as well as self-renewal capacity were evaluated by TUNEL, Live/Dead and colony assays, respectively. In the next step, to validate SVZ-NSCs identity in other species, a similar protocol was applied to isolate NSCs from adult rat's SVZ as well. Our findings revealed that isolated SVZ-NSCs from both monkey and rat preserve proliferation capacity in at least nine passages as confirmed by Ki67 expression. Additionally, both SVZ-NSCs sources are capable of self-renewal in addition to NESTIN, SOX2, and GFAP expression. The mortality was measured meager with over 95% viability according to TUNEL and Live/Dead assay results. Eventually, the multipotency of SVZ-NSCs appraised authentic after their differentiation into neurons, astrocytes, and oligodendrocytes. In this study, we proposed a reliable method for SVZ-NSCs in vitro maintenance and identification, which, we believe is a promising cell source for therapeutic approach to recover neurological disorders and injuries condition.
Subject(s)
Brain/metabolism , Cell Differentiation/genetics , Neural Stem Cells/metabolism , Neurons/metabolism , Adult Stem Cells/metabolism , Animals , Astrocytes/metabolism , Cell Proliferation/genetics , Cell Self Renewal/genetics , Haplorhini/genetics , Lateral Ventricles/metabolism , Neurogenesis/genetics , Oligodendroglia/metabolism , RatsABSTRACT
spinal cord injury (SCI) is a traumatic damage that can causes a loss of neurons around the lesion site and resulting in locomotor and sensory deficits. Currently, there is widely attempts in improvement of treatment strategy and cell delivering to the central nervous system (CNS). The usage of hyaluronic acid (HA), the main components of the ECM in CNS tissue and neural stem cells (NSCs) niche, is a good selection that can increase of viability and differentiation of NSCs. Importantly, we demonstrate that encapsulation of human embryonic stem cell derived-neural stem cells (hESC-NS) in HA-based hydrogel can increased differentiation these cells into oligodendrocytes and improved locomotor function.
Subject(s)
Cell Differentiation , Human Embryonic Stem Cells/cytology , Hyaluronic Acid , Neural Stem Cells/cytology , Regeneration , Stem Cell Transplantation , Animals , Cell Survival , Cells, Cultured , Disease Management , Fluorescent Antibody Technique , Human Embryonic Stem Cells/metabolism , Humans , Hydrogels , Male , Neural Stem Cells/metabolism , Rats , Spinal Cord Injuries/therapy , Tissue ScaffoldsABSTRACT
A large number of treatment approaches have been used for spinal cord injury improvement, a medically incurable disorder, and subsequently stem cell transplantation appears to be a promising strategy. The main objective of this study is to ascertain whether combinational therapy of human neural stem cells (hNSCs) together with lithium chloride improves cell survival, proliferation, and differentiation in a rat spinal contusion model, or not. Contusive spinal cord injury was implemented on Wistar male rats. Experimental groups comprised of: control, hNSCs transplanted, lithium chloride (Li), and hNSCs and lithium chloride (hNSCs + Li). In every experimental group, locomotor activity score and motor evoked potential (MEP) were performed to evaluate motor recovery as well as histological assessments to determine mechanisms of improvement. In accordance with our results, the hNSCs + Li and the Li groups showed significant improvement in locomotor scores and MEP. Also, Histological assessments revealed that transplanted hNSCs are capable of differentiation and migration along the spinal cord. Although NESTIN-positive cells were proliferated significantly in the Lithium group in comparison with control and the hNSCs + Li groups, the quantity of ED1 cells in the hNSCs + Li was significantly larger than the other two groups. Our results demonstrate that combinational therapy of hNSCs with lithium chloride and lithium chloride individually are adequate for ameliorating more than partial functional recovery and endogenous repair in spinal cord-injured rats.
