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Spinal cord injury (SCI) is a destructive condition that the cord can't send signals below the level of the injury. Despite advances in the medical and surgical care of SCI patients, no effective treatment exists for the neurological deficits of SCI. Cell therapy is a new approach for SCI, and preclinical models demonstrate that cell transplantation can improve some secondary events through neuroprotection and also restore lost tissue through regeneration. Neural Stem Cells (NSCs) are multipotent cells committed to the neural lineage that can self-renew. NSCs are found in both fetal and adult central nervous system (CNS). NSCs locate within specific niches in the adult CNS, including the subventricular zone in the lateral ventricles of the forebrain, the dentate gyrus of the hippocampus, olfactory bulb and the region of the central canal of the spinal cord. Transplantation of NSCs into injured tissue, promoted functional recovery with neuroprotective and neuroregenerative effects. Most studies with transplanted NSCs have shown modest recovery of the injured spinal cord. Adult mouse brain–derived NSCs transplanted into the injured rat spinal cord with concomitant infusion of growth factors promoted oligodendrocyte differentiation of the grafted NSCs, remyelination, and improved locomotor function. NSCs derived from fetal rat spinal cord differentiated into neurons that integrated into the injured cord and improved recovery, and transplanted NSCs combined with valproic acid administration promoted neuronal differentiation, resulting in restoration of disrupted neuronal circuitry and enhanced recovery. NSCs have also demonstrated some immunomodulatory and pathotropic ability by homing toward damaged tissue as well as secreting various neurotrophic factors and cytokines. Neuralstem cells also express nerve growth factors that are essential to the healthy function of the CNS. These could protect the patient’s own neurons from further degeneration due to injury. By solving some limitations in future, cell therapy can open a new window for treatment of SCI.

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It is estimated that annually 10 million people are affected by traumatic brain injury and it is one of the major causes of death and disability in accidents. Studies have shown the potential therapeutic value of neural stem cell therapies. Also, neural stem cell motility and migration to the site of injury has a great regeneration value of the damaged tissues. Extracellular and intracellular factors orchestrate this complicated process. In this work, we tried to elucidate the intracellular and indeed effectors of the cell motility and migration in neurosphere formations under in vitro conditions. After isolation and culture of bone marrow stromal cells (BMSCs) from rat the cells were cultured in DMEM/F12 medium supplemented with 2% B27, 20 ng/ml basic fibroblast growth factor, 20 ng/ml epidermal growth factor, 100 U/ml penicillin, and 100 mg/ml streptomycin. After passing the incubation time total RNA were extracted from the cells and cDNA synthesis were performed for different time i.e. at the times of 0, 1, 5 and 30 minutes as well as 1, 2, 4, 6, 12 and 24 hours. These cDNA were subjected to RT-PCR and real time RT-PCR reactions. At aforementioned different time courses RT-PCR and real time RT-PCR results showed that there are substantial differences in the expression of the genes which regulate polymerization and depolimerization of intracellular actin protein and thus cell cytoskeleton dynamics including Cdc42, Cttn, Pak1, Rock1 genes. Actin protein dynamic causes cell membrane protrusions and filopodia formation and thus cell migration. Discovering of the underlined signaling mechanisms and pathways that guide the cell motility has a great importance, especially neurosphere cell motility in the field of CNS regeneration medicine. In conclusion, our results show that Cdc42, Cttn, Pak1, Rock1 are effector genes in the cell motility of neurosphere formations.

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Spinal cord injury (SCI) is a serious condition that affects millions of people worldwide .The most causes for the SCI are motor vehicle accident (43%). Recent advances in stem cell research have brought closer the possibility of repairing the spinal cord. Cell therapy is an option in replacing the lost cells in the injured spinal cord. Adipose derived stem cells (ADSCs) are one of the kinds of stem cells that can be differentiation into neurosphere. Previous studies used the toxic factors and complicated methods. Here, we apply a nontoxic and efficient method of rat mesenchymal stem cells (MSCs) into neurosphere. As well as, the neurosphere can be differentiated in to glial and neural cells. Primary rat ADSCs were isolated from Wistar rats (200–300 g). Then MSCs derived ADSCs, cultured by DMEM medium supplemented with 10% fetal bovine serum. These cells evaluated by specific markers of MSCs and ADSCs such as CD49, CD90, CD105. By bioactive substance TNT then MSCs differentiated in to neurosphere in 4 groups that these groups compared with morphology. This differentiation do with nontoxic factor and by dose response of: 1 M, 0.1 M, 0.01 M and 0.25 M and time course at 72 hours. Diameter and number of this neurosphere evaluated every day in 4 groups. MSCs isolated from ADSCs then evaluated by immunocytochemistry that expressed CD90 (80%), CD49 (70%) fibronectin and negative marker CD45. Diameter and number of Neurosphere by 0.1 ng/ml was optimal dose for expansion. Cell therapy is an option in replacing the lost cells in the injured spinal cord. Source of patients need multiple delivery of cells in order to achieve the wanted results.

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Generation neural stem cells from neurosphere-derived adipose tissue using bioactive substance ATC. Adipose tissue from rat hypodermal and pararenal fat was digested with collagenase, followed by filter and centrifugation the isolated adipose stromal cells were cultured in dishes. These cells evaluated by specific markers of adipose-derived stem cells (ADSCs) such as bioactive substance ATC and then ADSCs differentionated in to neurosphere in four groups that compared morphologically. Diameter and number of this neurosphere evaluated every day in four groups. ADSCs are mesenchymal stem cells that can be extracted from adipose tissue and obtained by a less invasive method. ADSCs markers were measured by immunocytochemistery that expressed CD90 (80%), CD44 (70%) and fibronectin while CD45 didn’t express. Diameter and number of Neurosphere by (0.1 ng/ml) was optimal dose for expansion. Then these cells evaluated by neuroectodermal markers such as nestin and NF 68 that expressed>80% and this data approved by RT-PCR technique. This study develops a simplified, efficient, and nontoxic approach by lowest factors which derives a large number of neurospheres from Adipose-derived stem cells (ADSCs). With our newly devised approach 10 to 15 passage cells were used for in vitro differentiation. Neuronal differentiation was induced by incubation of the ADSCs with bioactive substance (ATC) induction media.

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Generation neural stem cells from neurosphere–derived bone marrow stem cells using bioactive substance ATC. Bone marrow cells (BMSCs) were isolated from rat. BMSCs cultured by DMEM/F12 medium supplemented with 10% fetal bovine serum. These cells evaluated by specific markers of BMSCs such as bioactive substance ATC, B27. Then BMSCs differentionated in to neurosphere and divided in two groups which were evaluated morphologically. Diameter and number of this neurosphere evaluated daily. BMSCs markers were measured by immunocytochemistery that expressed, CD 90 (75%) CD 44 (60%) fibronectin. Diameter and number of Neurosphere by 0.1 ng/ml was optimal dose for expansion. Then these cells evaluated by neuroectodermal markers such as nestin and NF 68, NF 200 and NF160, that expressed >80 % and this data approved by RT-PCR assay. This study develops a simplified, efficient, and nontoxic approach by lowest factors which derives a large number of neurospheres from BMSCs. With our newly devised approach 10 to 15 passage cells were used for in vitro differentiation. Neuronal differentiation was induced by incubation of the BMSCs with bioactive substance (ATC) induction media.

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In the last few years, a lot of preclinical studies showed the therapeutic potential of stem cells in spinal cord injury (SCI). Neural stem cells (NSCs) can be differentiated into all cell types of spinal cord. NSCs are cells that maintain the capacity to differentiate into brain-specific cell types, and may also replace or repair of injury in central nervous system. Here in, we describe the efficient conversion of Adipose derived stem cells (ADSCs) into a neural stem cell-like under the influence of a factor inducing non-toxic bioactive substance TNT. ADSCs were isolated from adipose tissue of Wistar rat and were cultured. ADSCs were treated with bioactive substance TNT. These cells grew in neurosphere-like structures and differentiated to neural stem cells. Immunocytochemistry and RT-PCR techniques were performed to evaluate early neuroectodermal markers including SOX2, OCT4, NANOG, NeuroD and nestin. Our results showed that these markers expressed in high level. Also, NSCs were immunoreactive to NF68, NF200 and nestin. In addition, we confirmed the expression of SOX2, OCT4, NANOG and NeuroD genes by RT-PCR assay. The findings of this study provide a new method to generate NSCs from ADSCs by using non-toxic bioactive substance TNT, which can be helpful in cell therapy of SCI and degenerative diseases.

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Spinal cord injury (SCI) is one of the devastating conditions leading to functional and neurological deficits following road traffic accidents. To date, there is no definite treatment for repairing damaged spinal cord tissue. In this regard, cell therapy opens a new window in front of scientists by using different cells such as mesenchymal stem cells, olfactory ensheathing cells, Schwann cells, neural stem cells and induced pluripotent stem cells. But, cell therapy faces some problems related to cell survival, migration and differentiation. Based on the results of recent studies, supporting the transplanted cells with a proper scaffold can be helpful. Among several scaffolds, hydrogels are outstanding one with several beneficial properties for neural tissue engineering. The future of SCI regeneration is probably linked to combinatorial approaches in which both cells and scaffolds play its significant roles.
 

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Spinal cord injury (SCI)-induced systemic inflammatory response affects multiple organs outside the spi­nal cord. Treatment options for such complications are lacking. Valproic acid (VPA) is a histone deacetylase inhibitor, acting directly at the level of gene transcription by inhibiting histone deacetylation and making transcription sites more accessible. Acetylation of histones is critical to cellular inflammatory and repair processes. A recent study demonstrated that VPA has effects on neuroprotection and neurogenesis for the treatment of the injured spinal cord. VPA can decreases glial apoptosis, neruoinflammation, neurotoxicity and autophagy during the secondary injury period, and upregulates prosurvival neurotrophic factors. The neuroprotective effects of VPA are interdepend and mediated by HDAC inhibition and GSK-3 inhibition. VPA increased several stages of neurogenesis, including the proliferation of endogenous neural stem cells, neuronal differentiation and maturation, neurite outgrowth, and synaptic integration. In addition, VPA can promote neurogenesis even after spinal cord cells are damaged, by controling the expression of important transcriptional factors and the activation of multiple signaling pathways. Furthermore, the effects and mechanisms of VPA on neuronal excitation mediated neuroprotection and neurogenesis are cooperated and interconnected in treating SCI. It is necessary to optimize VPA treatment processes for SCI on aspects of therapeutic timing, effective dosage, and reliable administration route. Combinatory strategies should be established to maximize the benefits of VPA and to reduce adverse events. Specific criteria must be met prior to translating VPA treatment for SCI from animal experiments to clinical trials.

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Spinal cord injury (SCI) is a destructive event that often lead to permanent neurologic deficit. Current clinical treatments are aimed at preventing secondary damage, promoting regeneration, and replacing destroyed spinal cord tissue, although effective treatments for SCI remain limited. Cell therapies for treating SCI are promising therapy for replacing dead cells, neuroprotection and axon regeneration. A number of different pluripotent, multipotent, and differentiating stem cells have been investigated so far for the treatment of SCI. Some of these cells have entered or will soon be entering clinical trials. Basic and pre-clinical experimental studies have highlighted the positive effects of Induced pluripotent stem cells (iPSCs) treatment after spinal cord and peripheral nerve injury. iPSCs are a type of pluripotent stem cell that directly can be generated from adult cells and their therapeutic effects are believed to be due to their potential to differentiate into neural precursor cells, neurons, oligodendrocytes, astrocytes  and neural crest cells that can act by replacing lost cells or providing environmental support. iPSCs can provide a cell source that has characteristics of embryonic stem cells. However, human iPSCs solve the ethical dilemma posed by human embryonic stem cells research. In addition, they can be sourced from autologous sources, which may decrease the risk of immune rejection.

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Spinal cord injury (SCI) is a devastating condition, with sudden loss of sensory, motor, and autonomic function distal to the level of trauma. The primary mechanical trauma causes necrosis, edema, hemorrhage, and vasospasm. A cascade of secondary pathophysiological mechanisms is induced, including ischemia, apoptosis, fluid and electrolyte disturbances, excitotoxicity, lipid peroxidation, production of free radicals, and an inflammatory response, resulting in further damage due to swelling and blood flow reduction. Cell therapy is a promising strategy for SCI, and preclinical models show that cell transplantation can improve some secondary events through neuroprotection and also restore lost tissue through regeneration. Neural stem/progenitor cells (NSPCs) are multipotent cells entrusted to the neural lineage that can self-renew and expanded In vitro. NSPCs are usually grown as free-floating neurospheres in serum-free medium supplemented with growth factors. It has been reported that neuronal differentiation of human fetal NSPC grafts after transplantation into the adult rat spinal cord. In addition, human fetal brain NSPCs transplanted into the contused cervical spinal cord produced significantly repair than controls.

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Epilepsy is neurological disorders that afflict many people around the world with a higher prevalence rate in children and in low income countries. Temporal lobe epilepsy (TLE) is result from hippocampal sclerosis is a neurological disorder with difficult treatment. Stem cells can transform into any type of cells such as glial cells, consequently stem cells can use for medical treatment. Stem cell therapy in epilepsy result in prophylaxis against epilepsy and improve cognitive function after seizures. Astrocytes   have many roles in the brain such as protection of neurons and endothelial cells, feeding, inhibiting over activation of microglia, modulate k changes, managing of extracellular ions, regulating density of y-amino butyric acid, glutamate and adenosine. Excessive activation of microglia cause brain inflammation that lead to epileptic seizures. Adult cell from patient have the capability to alter to embryonic cell and become stem cell by using transcription factors. Astrocytes by secretion of glial cell derived neurotrophic factor (GDNF), controlling the proliferation, adheration and movement of microglial cells .also astrocytes reduce generation of lipopolysaccharide (LPS), IL1B, TNF .astrocytes are as a source of protection mediators that decreased neuroinflammation. In this hypothesis I suggest using stem cell therapy in epilepsy to reduce neuroinflammation induced by microglia and reduce occurrence of seizures.

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Epilepsy is one of the most common neuroinflammatory disorders that affect more than 50 million people worldwide. Excessive electrical discharges in neurons following neural cell damage or loss leads to recurrent seizures, which are described as epilepsy. One of the most common and difficult to treat types of epilepsy is Temporal Lobe Epilepsy (TLE), which results from hippocampal sclerosis. Currently, drug therapy is one of the most used treatments for epilepsy, but anti-epileptic drugs can induce undesirable side effects and are not effective in all TLE patients. Therefore, developing new treatments for TLE is necessary. Recently, some studies have surveyed the use of stem cells for treatment of TLE. Stem cells have numerous significant advantages over current drug therapies for epilepsy. Researchers have used various stem cells in animal models for treatment of TLE, but there is no conclusive evidence in support of using stem cells for treating TLE yet. However it is important to acknowledge that this field is still in infancy, and the initial studies are promising. Thus, we suggest more researches need to be done on the use of stem cells for treatment of TLE.

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Neuroinflammation is a disorder that causes neurological disease. Neuroinflammation has a significant role in induce of Multiple sclerosis (MS) and one of the situations that must be treated stops the ongoing process of inflammation against the CNS by self-reactive lymphocytes. According to the successful results that were obtained from the pre-clinical phase of cell therapy, many studies were performed in clinical phase which resulted in the improvement of clinical symptoms, and in most of them, the quality of life and reduced relapsed were observed. In a comprehensive study, 500 MS patients worldwide were treated with Hematopoietic cells. 100% of the patients suppressed or reduced inflammation as well as the brain atrophy, which was also an inflammatory complication that was reduced within the patients. All studies in the field of cell therapy show high performance and effectiveness of this approach for the treatment of patients with multiple sclerosis. Even in the most severe stage of the disease (aggressive and resistant forms), the treatments resulted in a positive outcome within the patients; proving that this treatment is optimal for current patients suffering this disease. According to this successful therapeutic method, in recent years the complications were dropped and its severity was minimized crucially in patients who were under the age 40 and the duration of the disease was less than 5 years. Considering that these pharmacological interventions were available for MS patients to have numerous side effects such as Leukoencephalopathies, Thrombocytopenia, Autoimmune and Kidney disease, therefore Phase III clinical trials for comparing and selecting the best possible method is needed to enable the most effective diagnosis of cell therapy treatment for these patients. Also, cohort study in this field should be done to discover the advantages and disadvantages of this method.

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Neuroinflammation has a significant role in induce of Multiple sclerosis (MS) many approaches have been used to treat MS, but none of these methods have not been able to fully improve. One of the methods can suppress inflammation and regenerate the nervous system is the use of cell therapy. Using cell therapy in pre-clinic phase can be realized, it's mechanism and potency to suppress neuroinflammation. The best way that plenty of researchers use it to simulate the MS condition is experimental autoimmune encephalomyelitis (EAE) which is method can be induced neuroinflammation in laboratory animals. In this context, a lot of researches have been done on EAE model. Many of these studies have been done on mesenchymal stem cells (MSC). MSC is a heterogeneous subset of, mesoderm stromal progenitor cells that are almost derived from connective tissue. MSCs can be obtained from adipose tissue, bone marrow, and umbilical cord that regulatory and inhibitory effect on the immune system. Transplantation of adipose-derived stem cell (ASCs) has demonstrated striking therapeutic effects and unique immunomodulatory capacities when delivered at the peak or later in the course of the disease in EAE rats. Recent studies have shown that umbilical cord-derived mesenchymal stem cells (UC-MSCs) exert a regulatory effect on the functions of immune cells. UC-MSCs could improve the impaired function of T-regulator cells (Treg) from MS patients and also enhanced the capacity of Tregs to release IL-10. There is still controversy about the use of UC-MSCs and ASCs, and more research is needed to determine the advantages and the disadvantages of them also in vitro method such as EAE cannot simulate all condition of disease therefor more research in clinical phase should be done.

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