Induction of mice adult bone marrow mesenchymal stem cells into functional motor neuron-like cells (original) (raw)
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Differentiation of Mouse Bone-Marrow Mesenchymal Stem Cells into Motor Neuron Cells in vitro
Journal of Al-Nahrain University-Science, 2016
Motor neuron cell is responsible for transfer neural instruct from brain to peripheral muscle through spinal cord; therefore any defect in these cells or spinal cord will affect motion. This study was designed to induce differentiation of Bone marrow mesenchymal stem cells (BM-MSCs) into neuron cells. BM-MSCs were isolated from bone stroma of femur and tibia of albino male mice and tested immunocytochemically for CD44, CD90, and CD105 expression and showed positive staining, while showed negative staining for CD34. Differentiation of BM-MSCs to motor neuron involved two main steps. Firs; induction of BM-MSCs by addition of 1mM mercaptoethanol (BME) in fetal bovine serum (FBS) in minimum essential medium MEM for 24 h and 2mM BME in free serum media for 2h. In the second step of induction retinoic acid, sonic hedgehog and nerve growth factor were added in free serum MEM for 4 days. Results revealed that the differentiation medium used was very efficient in directing the BM-MSCs to motor neural cell and showed positive reactivity to specific motor neural markers that used for detection of motor neuron cells like microtubule associated protein-2 antibodies and acetylcholine transferase antibody.
Stem cell research & therapy, 2015
Severe spinal cord injury (SCI) often causes temporary or permanent damages in strength, sensation or autonomic functions below the site of the injury. So far there is still no effective treatment for SCI. Mesenchymal stem cells (MSCs) have been used to repair injured spinal cord as an effective strategy. However, the low neural differentiation frequency of MSCs has limited its application. The present study attempted to explore whether the grafted MSC-derived neural-like cells in a gelatin sponge (GS) scaffold could maintain neural features or transdifferentiate into myelin-forming cells in the spinal cord transected. We constructed an engineered tissue by co-seeding of MSCs with genetically enhanced expression of neurotrophin-3 (NT-3) and its high affinity receptor tropomyosin receptor kinase C (TrkC), respectively, into a 3-dimensional GS scaffold to promote the MSCs differentiating into neural-like cells, and transplanted it into the gap of rat spinal cord transected completely....
Mesenchymal stem cells as an alternative for Schwann cells in rat spinal cord injury
Iranian biomedical journal, 2013
Spinal cord has a limited capacity to repair; therefore, medical interventions are necessary for treatment of injuries. Transplantation of Schwann cells has shown a great promising result for spinal cord injury (SCI). However, harvesting Schwann cell has been limited due to donor morbidity and limited expansion capacity. Furthermore, accessible sources such as bone marrow stem cells have drawn attentions to themselves. Therefore, this study was designed to evaluate the effect of bone marrow-derived Schwann cell on functional recovery in adult rats after injury. Mesenchymal stem cells were cultured from adult rats' bone marrow and induced into Schwann cells in vitro. Differentiation was confirmed by immunocytochemistry and RT-PCR. Next, Schwann cells were seeded into collagen scaffolds and engrafted in 3 mm lateral hemisection defects. For 8 weeks, motor and sensory improvements were assessed by open field locomotor scale, narrow beam, and tail flick tests. Afterwards, lesioned s...
Neuroscience Letters, 2009
Mesenchymal stem cells (MSCs) have demonstrated a measurable therapeutic effect following transplantation into animal models of spinal cord injury. However, the mechanism(s) by which transplanted cells promote nerve regeneration and/or functional recovery remains indeterminate. Several studies have suggested that MSCs promote tissue repair via secretion of trophic factors, but delineating the effect of such factors is difficult due to the complexity of the in vivo systems. Therefore, we developed an organotypic spinal cord slice culture system that can be sustained for sufficient periods of time in vitro to evaluate nerve regeneration as an ex vivo model of spinal cord injury. Using this model, we demonstrate that treatment of lumbar slices of spinal cord with lysolecithin induced a significant degree of cell death and demyelination of nerve fibers, but that these effects were ameliorated to a significant extent following co-culture of slices with human MSCs (hMSCs). The results indicate that transplanted hMSCs alter the tissue microenvironment in a way that promotes survival of endogenous cells, including injured neurons, immature oligodendrocytes and oligodendrocyte progenitor cells. This ex vivo culture system represents a useful tool to further dissect the mechanism(s) by which MSCs promote regeneration of injured nervous tissue.
PLoS ONE, 2013
Spinal cord injury triggers irreversible loss of motor and sensory functions. Numerous strategies aiming at repairing the injured spinal cord have been studied. Among them, the use of bone marrow-derived mesenchymal stem cells (BMSCs) is promising. Indeed, these cells possess interesting properties to modulate CNS environment and allow axon regeneration and functional recovery. Unfortunately, BMSC survival and differentiation within the host spinal cord remain poor, and these cells have been found to have various adverse effects when grafted in other pathological contexts. Moreover, paracrinemediated actions have been proposed to explain the beneficial effects of BMSC transplantation after spinal cord injury. We thus decided to deliver BMSC-released factors to spinal cord injured rats and to study, in parallel, their properties in vitro. We show that, in vitro, BMSC-conditioned medium (BMSC-CM) protects neurons from apoptosis, activates macrophages and is pro-angiogenic. In vivo, BMSC-CM administered after spinal cord contusion improves motor recovery. Histological analysis confirms the pro-angiogenic action of BMSC-CM, as well as a tissue protection effect. Finally, the characterization of BMSC-CM by cytokine array and ELISA identified trophic factors as well as cytokines likely involved in the beneficial observed effects. In conclusion, our results support the paracrine-mediated mode of action of BMSCs and raise the possibility to develop a cell-free therapeutic approach.
Cellular and Molecular Neurobiology, 2006
Human mesenchymal stem cells (hMSCs) derived from adult bone marrow represent a potentially useful source of cells for cell replacement therapy after nervous tissue damage. They can be expanded in culture and reintroduced into patients as autografts or allografts with unique immunologic properties. The aim of the present study was to investigate (i) survival, migration, differentiation properties of hMSCs transplanted into non-immunosuppressed rats after spinal cord injury (SCI) and (ii) impact of hMSC transplantation on functional recovery. Seven days after SCI, rats received i.v. injection of hMSCs (2 × 10 6 in 0.5 mL DMEM) isolated from adult healthy donors. Functional recovery was assessed by Basso-Beattie-Bresnahan (BBB) score weekly for 28 days. Our results showed gradual improvement of locomotor function in transplanted rats with statistically significant differences at 21 and 28 days. Immunocytochemical analysis using human nuclei (NUMA) and BrdU antibodies confirmed survival and migration of hMSCs into the injury site. Transplanted cells were found to infiltrate mainly into the ventrolateral white matter tracts, spreading also to adjacent segments located rostro-caudaly to the injury epicenter. In double-stained preparations, hMSCs were found to differentiate into oligodendrocytes (APC), but not into cells expressing neuronal markers (NeuN). Accumulation of GAP-43 regrowing axons within damaged white matter tracts after transplantation was observed.
Experimental Neurology, 2013
Neurotrophins and the transplantation of bone marrow-derived stromal cells (MSCs) are both candidate therapies targeting spinal cord injury (SCI). While some studies have suggested the ability of MSCs to transdifferentiate into neural cells, other SCI studies have proposed anti-inflammatory and other mechanisms underlying established beneficial effects. We grafted rat MSCs genetically modified to express MNTS1, a multineurotrophin that binds TrkA, TrkB and TrkC, and p75 NTR receptors or MSC-MNTS1/p75 − that binds mainly to the Trk receptors. Seven days after contusive SCI, PBS-only, GFP-MSC, MSC-MNTS1/GFP or MSC-MNTS1/p75 − /GFP were delivered into the injury epicenter. All transplanted groups showed reduced inflammation and cystic cavity size compared to control SCI rats. Interestingly, transplantation of the MSC-MNTS1 and MSC-MNTS1/p75 − , but not the naïve MSCs, enhanced axonal growth and significantly prevented cutaneous hypersensitivity after SCI. Moreover, transplantation of MSC-MNTS1/p75 − promoted angiogenesis and modified glial scar formation. These findings suggest that MSCs transduced with a multineurotrophin are effective in promoting cell growth and improving sensory function after SCI. These novel data also provide insight into the neurotrophin-receptor dependent mechanisms through which cellular transplantation leads to functional improvement after experimental SCI.
A comparative study of three different types of stem cells for treatment of rat spinal cord injury
Cell transplantation, 2016
Three different sources of human stem cells – bone marrow mesenchymal stem cells (MSCs), and two neural progenitors (NPs) derived from immortalized spinal fetal cell line (SPC 01), or induced pluripotent stem cells (iPS-NPs) – were compared in the treatment of a balloon-induced spinal cord compression lesion in rats. One week after lesioning, the rats received either MSCs (intrathecally) or NPs (SPC-01 cells, or iPS-NPs, intraspinally), or saline. The rats were assessed for their locomotor skills (BBB, flat beam test, rotarod). Morphometric analyses of spared white and grey matter, axonal sprouting and glial scar formation, as well as qPCR and Luminex assay were conducted to detect endogenous gene expression, while inflammatory cytokine levels were performed to evaluate the host tissue response to stem cell therapy. The highest locomotor recovery was observed in iPS-NP-grafted animals, which also displayed the highest amount of preserved white and grey matter. Grafted iP...