Effect of GDNF on Morphology, Proliferation, and Phagocytic Activity of Rat Neonatal Cortex Isolated Microglia

© 2016, Springer Science+Business Media New York.Microglia are the main defenders of the central nervous system and at the same time are involved in the pathogenesis of various neurological disorders. Microglia hyperactivity or phagocytic impairment exacerbates degenerative processes in nervous tissue leading to further loss of function. A variety of factors and cytokines may modify microglia function. In our study, it was shown that glial cell line-derived neurotrophic factor (GDNF), a well-known neuroprotective molecule, decreases phagocytic activity of microglia in vitro model of spinal cord injury. Recombinant adenovirus encoding GDNF (Ad5-GDNF) transfected microglia have shown the same effect and can be potentially used as a therapeutic agent in case of neurotrauma due to its debris phagocytic and GDNF-associated neuroprotective role

Resistência à seca em seringueira. II. Crescimento e partição de assimilados em clones submetidos a déficit hídrico

Plants from IAN 717, IAN 873, IAN 2903, IAN 3087, IAN 6323 and Fx 3899 rubber tree (Hevea spp.) clones grown in a greenhouse were subjected to cycles of water deficit with the objective of evaluating the effects on growth and assimilation partitioning. After 185 days and five cycles of stress, a reduction in the number of leaves, in the number of flushes, and in the length and diameter of the shoots occurred in the majority of the clones. The IAN 2903 progeny showed an increase in the number of leaves (4.7%) and in the number of flushes (10%). Shoots of the IAN 717 and IAN 3087 progenies were the least affected in their growth. The assimilation partitioning showed a modification in relation to the sinks occurring with the deficit; the prefered sinks were the root in the Fx 3899 and 873 progenies the stem in the IAN 3087 and IAN 6323 progenies and the leaf in the IAN 717 and IAN 2903 progenies. The ratio of the aerial part to the root system was greater in the IAN 2903 progeny and smaller in the Fx 3899 progeny. Under water stress no significant changes occurred in the specific leaf area production values for leaf area and in the leaf area ratio, but the net assimilation value and the accumulation and production of dry matter decreased differentially when submitted to water deficit. The clone least affected by the stress was IAN 3087. The greatest reductions on studied parameters occurred in Fx 3899. The results observed in each clone were discussed in relation to each parameter studied.Plantas de seringueira (Hevea spp) oriundas dos clones IAN 717, IAN 873, IAN 2903, IAN 3087. IAN 6323 e Fx 3899 foram cultivadas em casa-de-vegetação e submetidas a ciclos de estresse hídrico, com objetivo de avaliar comparativamente os efeitos sobre o crescimento e a partição de assimilados. Após 185 dias e cinco ciclos de estresse, o número de folhas, número de lançamentos, comprimento e diâmetro das brotações na maioria dos clones diminuíram. O IAN 2903 aumentou o número de folhas (4,7%) e o de lançamento (10%). As brotações do IAN 717 e IAN 3087 foram as menos afetadas no seu crescimento. A partição de assimilados mostrou uma modificação na preferência dos drenos com o déficit, sendo que alocação preferencial para a raiz foi do Fx 3899 e IAN 873, para o caule do IAN 3087 e IAN 6323 e para a folha do IAN 717 e IAN 2903. A relação parte aérea/sistema radicular foi maior no IAN 2903 e menor no Fx 3899. O déficit não provocou mudanças significativas na área foliar específica e na razão de peso foliar. Entretanto, a área foliar total, taxa de produção de área foliar, razão de área foliar, taxa assimilatória líquida, acúmulo e produção de matéria seca decresceram diferencialmente quando submetidos à seca. O clone menos afetado pelo estresse foi o IAN 3087. As maiores reduções nos parâmetros estudados ocorreram no Fx 3899. São discutidos os resultados de cada clone com relação a cada parâmetro estudado

Human umbilical cord blood cell transplantation in neuroregenerative strategies

© 2017 Galieva, Mukhamedshina, Arkhipova and Rizvanov. At present there is no effective treatment of pathologies associated with the death of neurons and glial cells which take place as a result of physical trauma or ischemic lesions of the nervous system. Thus, researchers have high hopes for a treatment based on the use of stem cells (SC), which are potentially able to replace dead cells and synthesize neurotrophic factors and other molecules that stimulate neuroregeneration. We are often faced with ethical issues when selecting a source of SC. In addition to precluding these, human umbilical cord blood (hUCB) presents a number of advantages when compared with other sources of SC. In this review, we consider the key characteristics of hUCB, the results of various studies focused on the treatment of neurodegenerative diseases (Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis), ischemic (stroke) and traumatic injuries of the nervous system and the molecular mechanisms of hUCB-derived mononuclear and stem cells

Effects of transplantation of human cord blood mononuclear cells expressing the recombinant VEGF and FGF2 genes into spinal cord traumatic injury sites in rats

A model of dosed TVIII spinal cord contusion trauma in rats was used to study the effects of immediate single-dose transplantation of human cord blood mononuclear cells transformed with the recombinant genes for neurotrophic factors - vascular endothelia growth factor (VEGF) and fibroblast growth factor 2 (FGF2) - into the injury zone. A further group of animals, in the same conditions, received the same cells transfected with plasmid pEGFP-N2. EGFP-labeled cells were detected in the white matter for 21 days after transplantation at distances of at least 10 mm in the rostral and caudal directions from the administration point. By 30 days after transplantation with cells transfected with plasmid pBud-VEGF-FGF2, the area of intact gray matter 3 mm from the trauma epicenter increased by more than 60%. By this time, the outer areas of the white matter in animals of this group, 1.5 cm from the trauma epicenter, showed an average 30% increase in the number of perivascular cells expressing platelet-derived growth factor β receptors (PDGFβR). Addition of therapeutic genes VEGF an FGF2 to the trauma injury zone and their expression in carrier cells stimulated vascularization and post-traumatic regeneration of the spinal cord. © 2013 Springer Science+Business Media New York

Usage of plasmid vector carrying vegf and fgf2 genes after spinal cord injury in rats

Using rat model of spinal cord contusion injury at TVIII, we compared the effectiveness of immediate single transplantation of human mononuclear umbilical cord blood cells transfected with pBud-VEGF-FGF2 plasmid and immediate direct injection of the same plasmid into the lesion area. The results suggest that the delivery of therapeutic genes vegf and fgf2 in cells is more effective than direct injection of plasmid DNA with the same genes (judging from the number of myelinated fibers). Better tissue preservation and motor function recovery in experiments with direct injection of plasmid pBud-VEGF-FGF2 suggest that direct gene therapy seems to be an effective additional procedure to the method of gene delivery with transfected stem and progenitor cells. © 2013 Springer Science+Business Media New York

Post-Traumatic Changes in the Spinal Cord in Rats after Transplantation of Mononuclear Cells from Human Umbilical Blood Modified with the vegf and fgf2 Genes

Experiments were carried out using 25 white laboratory rats. A model of dosed contusional trauma to the spinal cord at the T VIII level in rats was used to measure the areas of pathological cavities and the numbers of myelinated fibers in the outer zones of the white matter were counted after single rapid doses of human umbilical blood mononuclear cells transfected with a plasmid carrying the vegf and fgf2 genes into the injured area. Animals of the control group received the same cells in analogous conditions, but transfected with plasmid pEGFP-N2, carrying the gene for enhanced green fluorescent protein (egfp). By postadministration day 30, the total area of pathological cavities in the outer zones of the white matter on transverse sections of the spinal cord 3 mm from the trauma epicenter in the caudal direction was more than two times smaller in animals of the experimental group than in controls. The numbers of myelinated fibers in the same white matter zones at the same distance from the trauma epicenter in the caudal and rostral directions were an average of 20% greater than in controls, while at a distance of 5 mm in the rostral direction the number was 40-70% greater. Administration of the therapeutic genes vegf and fgf2 into the injured area decreased cavitation, restricted the processes of secondary degeneration and maintained the number of myelinated fibers in the damaged spinal cord. © 2012 Springer Science+Business Media New York

Adenoviral vector carrying glial cell-derived neurotrophic factor for direct gene therapy in comparison with human umbilical cord blood cell-mediated therapy of spinal cord injury in rat

© 2015 International Spinal Cord Society Study design:Experimental study.Objective:To evaluate the treatment of spinal cord injury with glial cell-derived neurotrophic factor (GDNF) delivered using an adenoviral vector (AdV-GDNF group) in comparison with treatment performed using human umbilical cord blood mononuclear cells (UCB-MCs)-transduced with an adenoviral vector carrying the GDNF gene (UCB-MCs+AdV-GDNF group) in rat.Setting:Kazan, Russian Federation.Methods:We examined the efficacy of AdV-GDNF and UCB-MCs+AdV-GDNF therapy by conducting behavioral tests on the animals and morphometric studies on the spinal cord, performing immunofluorescence analyses on glial cells, investigating the survival and migration potential of UCB-MCs, and evaluating the expression of the recombinant GDNF gene.Results:At the 30th postoperative day, equal positive locomotor recovery was observed after both direct and cell-based GDNF therapy. However, after UCB-MCs-mediated GDNF therapy, the area of preserved tissue and the number of spared myelinated fibers were higher than those measured after direct GDNF gene therapy. Moreover, we observed distinct changes in the populations of glial cells; expression patterns of the specific markers for astrocytes (GFAP, S100B and AQP4), oligodendrocytes (PDGFαR and Cx47) and Schwann cells (P0) differed in various areas of the spinal cord of rats treated with AdV-GDNF and UCB-MCs+AdV-GDNF.Conclusion:The differences detected in the AdV-GDNF and UCB-MCs+AdV-GDNF groups could be partially explained by the action of UCB-MCs. We discuss the insufficiency and the advantages of these two methods of GDNF gene delivery into the spinal cord after traumatic injury.Spinal Cord advance online publication, 29 September 2015; doi:10.1038/sc.2015.161

Adenoviral vector carrying glial cell-derived neurotrophic factor for direct gene therapy in comparison with human umbilical cord blood cell-mediated therapy of spinal cord injury in rat

© 2016 International Spinal Cord Society.Study design:Experimental study.Objective:To evaluate the treatment of spinal cord injury with glial cell-derived neurotrophic factor (GDNF) delivered using an adenoviral vector (AdV-GDNF group) in comparison with treatment performed using human umbilical cord blood mononuclear cells (UCB-MCs)-transduced with an adenoviral vector carrying the GDNF gene (UCB-MCs+AdV-GDNF group) in rat.Setting:Kazan, Russian Federation.Methods:We examined the efficacy of AdV-GDNF and UCB-MCs+AdV-GDNF therapy by conducting behavioral tests on the animals and morphometric studies on the spinal cord, performing immunofluorescence analyses on glial cells, investigating the survival and migration potential of UCB-MCs, and evaluating the expression of the recombinant GDNF gene.Results:At the 30th postoperative day, equal positive locomotor recovery was observed after both direct and cell-based GDNF therapy. However, after UCB-MCs-mediated GDNF therapy, the area of preserved tissue and the number of spared myelinated fibers were higher than those measured after direct GDNF gene therapy. Moreover, we observed distinct changes in the populations of glial cells; expression patterns of the specific markers for astrocytes (GFAP, S100B and AQP4), oligodendrocytes (PDGFαR and Cx47) and Schwann cells (P0) differed in various areas of the spinal cord of rats treated with AdV-GDNF and UCB-MCs+AdV-GDNF.Conclusion:The differences detected in the AdV-GDNF and UCB-MCs+AdV-GDNF groups could be partially explained by the action of UCB-MCs. We discuss the insufficiency and the advantages of these two methods of GDNF gene delivery into the spinal cord after traumatic injury

Intravenous Transplantation of Human Umbilical Cord Blood Mononuclear Cells Overexpressing Nerve Growth Factor Improves Spatial Memory in APP/PS1 Transgenic Mice with a Model of Alzheimer’s Disease

© 2017, Springer Science+Business Media, LLC, part of Springer Nature. Alzheimer’s disease is a progressive incurable neurodegenerative disease manifested by dementia and other cognitive disorders. Gene-cell therapy is one of the most promising trends in the development of treatment for Alzheimer’s disease. The study was aimed to evaluate the therapeutic potential of intravenous transplantation of human umbilical cord blood mononuclear cells (UCBMCs) transduced with adenoviral vectors overexpressing nerve growth factor (NGF) for the treatment of Alzheimer’s disease in an APP/PS1 transgenic mice model. The transplantation of NGF-expressing UCBMCs was found to improve spatial memory and decrease anxiety in APP/PS1 mice. Grafted cells and their expression of NGF were detected in the cortex and hippocampus of transgenic mice in the period up to 90 days after transplantation. Thus, gene-cell therapy based on the use of NGF-overexpressing UCBMCs is a promising approach for the development of Alzheimer’s disease treatments

Assessment of glial scar, tissue sparing, behavioral recovery and axonal regeneration following acute transplantation of genetically modified human umbilical cord blood cells in a rat model of spinal cord contusion

©2016 Mukhamedshina et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Objective and Methods This study investigated the potential for protective effects of human umbilical cord blood mononuclear cells (UCB-MCs) genetically modified with the VEGF and GNDF genes on contusion spinal cord injury (SCI) in rats. An adenoviral vector was constructed for targeted delivery of VEGF and GDNF to UCB-MCs. Using a rat contusion SCI model we examined the efficacy of the construct on tissue sparing, glial scar severity, the extent of axonal regeneration, recovery of motor function, and analyzed the expression of the recombinant genes VEGF and GNDF in vitro and in vivo. Results Transplantation of UCB-MCs transduced with adenoviral vectors expressing VEGF and GDNF at the site of SCI induced tissue sparing, behavioral recovery and axonal regeneration comparing to the other constructs tested. The adenovirus encoding VEGF and GDNF for transduction of UCB-MCs was shown to be an effective and stable vehicle for these cells in vivo following the transplantation into the contused spinal cord. Conclusion Our results show that a gene delivery using UCB-MCs-expressing VEGF and GNDF genes improved both structural and functional parameters after SCI. Further histological and behavioral studies, especially at later time points, in animals with SCI after transplantation of genetically modified UCB-MCs (overexpressing VEGF and GDNF genes) will provide additional insight into therapeutic potential of such cells