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Role of Sympathic Nervous System and RGM-A in the Resolution of Inflammation

ABSTRACT: Protein and phosphorylation (Phospho Explorer Antibody Array, FullMoonBioscience, #PEX100) profiling of peritoneal monocytes (pooled lavages from 3-4 mice / condition) was carried out according to the manufacturer’s instructions.

ORGANISM(S): Mus musculus Rattus norvegicus Homo sapiens

PROVIDER: GSE122569 | GEO | 2018/12/19

REPOSITORIES: GEO

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Project description:It has been widely accepted that mammalian neural stem cells (NSCs) only exist in central nervous system (CNS). Here, we challenge this concept and show that peripheral NSCs (pNSCs) could be isolated out of CNS from mouse embryonic limbs, postnatal tail and adult lung tissues. Derived-pNSCs express multiple NSC-specific markers, exhibit cell morphology, self-renewing capacity, genome-wide transcriptional profile, epigenetic features similar to those of brain control NSCs. Derived-pNSCs can differentiate into astrocytes, electrophysiologically functional neurons, and oligodendrocytes in vitro and in vivo, indicating multipotency. Lineage-tracing results reveal that pNSCs are progeny cells of neural epithelial cells, but not neural crest cells. Our finding of pNSCs out of CNS expands the field of developmental neuroscience and presents an alternative potential strategy for neural regenerative therapy. Generation of pNSCs: To generate pNSCs, embryonic limbs cells, adult lung cells and postnatal tail cells were cultured in simple defined NSC medium: DMEM/F-12 [1:1] with 2% B27 w/o Vitamin A, 1% N2 (both Gibco), 1x Glutmax (Gibco), 1x penicillin-streptomycin (Sigma), supplemented with 10 ng/ml EGF and 10 ng/ml human bFGF (both from Invitrogen). Medium was changed every 24h. After pNSCs clusters were observed, each single cluster colony was manually picked and cultured in 96 well individually. Microarray and data analysis: RNA samples to be analysed by microarrays were prepared using Qiagen RNeasy columns with on-column DNA digestion. 300 ng of total RNA per sample was used as input into a linear amplification protocol (Ambion) that involved synthesis of T7-linked double-stranded cDNA and 12 h of in vitro transcription incorporating biotin-labelled nucleotides. Purified and labelled cRNA was then hybridized for 18 h onto MouseRef-8 v2 expression BeadChips (Illumina) following the manufacturer’s instructions. After washing as recommended, chips were stained with streptavidin-Cy3 (GE Healthcare) and scanned using the iScan reader (Illumina) and accompanying software. Samples were exclusively hybridized as biological replicates. The bead intensities were mapped to gene information using BeadStudio 3.2 (Illumina). Background correction was performed using the Affymetrix Robust Multi-array Analysis (RMA) background correction model. Variance stabilization was performed using the log2 scaling and gene expression normalization was calculated with the method implemented in the lumi package of R-Bioconductor.

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Role of Sympathic Nervous System and RGM-A in the Resolution of Inflammation

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