In vivo tracking of human neural progenitor cells in the rat brain using bioluminescence imaging

AbstractBackgroundStem cell therapies appear promising for treating certain neurodegenerative disorders and molecular imaging methods that track these cells in vivo could answer some key questions regarding their survival and migration. Bioluminescence imaging (BLI), which relies on luciferase expression in these cells, has been used for this purpose due to its high sensitivity.New methodIn this study, we employ BLI to track luciferase-expressing human neural progenitor cells (hNPCLuc2) in the rat striatum long-term.ResultsWe show that hNPCLuc2 are detectable in the rat striatum. Furthermore, we demonstrate that using this tracking method, surviving grafts can be detected in vivo for up to 12 weeks, while those that were rejected do not produce bioluminescence signal. We also demonstrate the ability to discern hNPCLuc2 contralateral migration.Comparison with existing methodsSome of the advantages of BLI compared to other imaging methods used to track progenitor/stem cells include its sensitivity and specificity, low background signal and ability to distinguish surviving grafts from rejected ones over the long term while the blood–brain barrier remains intact.ConclusionsThese new findings may be useful in future preclinical applications developing cell-based treatments for neurodegenerative disorders

Additional file 5: Table E4. of RNA-sequencing analysis of lung primary fibroblast response to eosinophil-degranulation products predicts downstream effects on inflammation, tissue remodeling and lipid metabolism

Upstream Regulators of the genes listed in our dataset (target genes, Additional file 3: Table E2 and Additional file 4: Table E3) as predicted by IPA analysis. (PDF 141 kb

La Charente

31 octobre 18791879/10/31 (A8,N2098)-1879/10/31.Appartient à l’ensemble documentaire : PoitouCh

Additional file 9: Figure E3. of RNA-sequencing analysis of lung primary fibroblast response to eosinophil-degranulation products predicts downstream effects on inflammation, tissue remodeling and lipid metabolism

Global Molecular Network #2 generated by IPA downstream analysis. Interactions between the genes of our dataset #1 (300 genes) in relation with other neighboring genes present in a Global Molecular Network. Thirty-five total genes or group of genes are present in each network. This network characterized genes related to cellular movement, hematological system development and function and immune cell trafficking. It includes 15 genes and 4 groups from our dataset. (PDF 332 kb

Synergistic Effects of GDNF and VEGF on Lifespan and Disease Progression in a Familial ALS Rat Model

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the progressive loss of motor neurons in the brain and spinal cord. We have recently shown that human mesenchymal stem cells (hMSCs) modified to release glial cell line-derived neurotrophic factor (GDNF) decrease disease progression in a rat model of ALS when delivered to skeletal muscle. In the current study, we determined whether or not this effect could be enhanced by delivering GDNF in concert with other trophic factors. hMSC engineered to secrete GDNF (hMSC-GDNF), vascular endothelial growth factor (hMSC-VEGF), insulin-like growth factor-I (hMSC-IGF-I), or brain-derived neurotrophic factor (hMSC-BDNF), were prepared and transplanted bilaterally into three muscle groups. hMSC-GDNF and hMSC-VEGF prolonged survival and slowed the loss of motor function, but hMSC-IGF-I and hMSC-BDNF did not have any effect. We then tested the efficacy of a combined ex vivo delivery of GDNF and VEGF in extending survival and protecting neuromuscular junctions (NMJs) and motor neurons. Interestingly, the combined delivery of these neurotrophic factors showed a strong synergistic effect. These studies further support ex vivo gene therapy approaches for ALS that target skeletal muscle

Autophagy Protects against Eosinophil Cytolysis and Release of DNA

The presence of eosinophils in the airway is associated with asthma severity and risk of exacerbations. Eosinophils deposit their damaging products in airway tissue, likely by degranulation and cytolysis. We previously showed that priming blood eosinophils with IL3 strongly increased their cytolysis on aggregated IgG. Conversely, IL5 priming did not result in significant eosinophil cytolysis in the same condition. Therefore, to identify critical events protecting eosinophils from cell cytolysis, we examined the differential intracellular events between IL5- and IL3-primed eosinophils interacting with IgG. We showed that both IL3 and IL5 priming increased the eosinophil adhesion to IgG, phosphorylation of p38, and production of reactive oxygen species (ROS), and decreased the phosphorylation of cofilin. However, autophagic flux as measured by the quantification of SQSTM1-p62 and lipidated-MAP1L3CB over time on IgG, with or without bafilomycin-A1, was higher in IL5-primed compared to IL3-primed eosinophils. In addition, treatment with bafilomycin-A1, an inhibitor of granule acidification and autophagolysosome formation, enhanced eosinophil cytolysis and DNA trap formation in IL5-primed eosinophils. Therefore, this study suggests that increased autophagy in eosinophils protects from cytolysis and the release of DNA, and thus limits the discharge of damaging intracellular eosinophilic contents

Endogenous Semaphorin-7A Impedes Human Lung Fibroblast Differentiation

<div><p>Semaphorin-7A is a glycosylphosphatidylinositol-anchored protein, initially characterized as an axon guidance protein. Semaphorin-7A also contributes to immune cell regulation and may be an essential pro-fibrotic factor when expressed by non-fibroblast cell types (exogenous). In mouse models, semaphorin-7A was shown to be important for TGF-ß1-induced pulmonary fibrosis characterized by myofibroblast accumulation and extracellular matrix deposition, but the cell-specific role of semaphorin-7A was not examined in fibroblasts. The purpose of this study is to determine semaphorin-7A expression by fibroblasts and to investigate the function of endogenously expressed semaphorin-7A in primary human lung fibroblasts (HLF).</p><p>Herein, we show that non-fibrotic HLF expressed high levels of cell surface semaphorin-7A with little dependence on the percentage of serum or recombinant TGF-ß1. Semaphorin-7A siRNA strongly decreased semaphorin-7A mRNA expression and reduced cell surface semaphorin-7A. Reduction of semaphorin-7A induced increased proliferation and migration of non-fibrotic HLF. Also, independent of the presence of TGF-ß1, the decline of semaphorin-7A by siRNA was associated with increased α-smooth muscle actin production and gene expression of periostin, fibronectin, laminin, and serum response factor (SRF), indicating differentiation into a myofibroblast. Conversely, overexpression of semaphorin-7A in the NIH3T3 fibroblast cell line reduced the production of pro-fibrotic markers. The inverse association between semaphorin-7A and pro-fibrotic fibroblast markers was further analyzed using HLF from idiopathic pulmonary fibrosis (IPF) (n = 6) and non-fibrotic (n = 7) lungs. Using these 13 fibroblast lines, we observed that semaphorin-7A and periostin expression were inversely correlated. In conclusion, our study indicates that endogenous semaphorin-7A in HLF plays a role in maintaining fibroblast homeostasis by preventing up-regulation of pro-fibrotic genes. Therefore, endogenous and exogenous semaphorin-7A may have opposite effects on the fibroblast phenotype.</p></div

Semaphorin-7A expression is inversely correlated with periostin expression in HLF.

<p>Expression of semaphorin-7A (sema7A), periostin, fibronectin and collagen-1 (COL1A1) were measured by real-time PCR in fibroblasts from non-fibrotic (●, n = 7) and IPF lung (■, n = 6). Semaphorin-7A expression was inversely associated with periostin expression level (<i>r</i>p = -0.608, <i>p</i> = 0.027, n = 13).</p

Semaphorin-7A diminishes HLF proliferation and migration.

<p>HLF derived from non-fibrotic lungs were treated with either control or semaphorin-7A siRNA and cultured in 1% or 10% FBS for 48 h. A/ BrdU was incubated with cells for 6 h. Absorbance was measured using a spectrophotometric plate reader at dual wavelengths of 450–550 nm. Each condition was performed in quadruplicate (4 wells) and graphs show a mean ± SEM of 3 experiments. B/ Cells were resuspended in 0.1% FBS and migration on plastic or plexin-C1 (10 μg/ml) toward medium with 10% FBS was measured after 20 h. Graph shows an average ± SEM of 4 experiments, and * indicates statistical difference between control-siRNA and sema7A-siRNA-treated HLF.</p