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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.
2023-06-02 | GSE151649 | GEO
Project description:It has been widely accepted that mammalian neural stem cells (NSCs) exist only in the central nervous system. Here, we report that peripheral NSCs (pNSCs) exist outside of the CNS and can be isolated—from mouse embryonic limb, postnatal lung, tail, dorsal root ganglia, and adult lung tissues. pNSCs are from Sox1+ cells and originate from neuroepithelial cells. Many pNSCs that migrate out of the neural tube can differentiate into neurons and limited glia cells during embryo development. We found the existence of pNSCs provides previously unidentified insight of the mammalian nervous system development and presents an alternative potential strategy for neural regenerative therapy. Our study demonstrates that directly converted cells can be a promising source for generating transplantable assembloids. Mice and Cells: RNA-seq : RNA quality was verified on an Agilent Bioanalyzer Nano Eukaryote chip. 1 mg of total RNA was used for poly-A selection with NEBNext PolyA mRNA magnetic isolation module and subsequent cDNA synthesis, Illumina Tru-seq adaptor ligation and library preparation were performed with NEBNext Ultra RNA Library Prep Kit (NEB) according to the manufacturer’s instructions. Library quality and concentration were determined using an Agilent Bioanalyzer DNA1000 chip and a Qubit fluorometer. Libraries were sequenced on Illumina NextSeq500 in single-end mode. HISAT24 was used to align the RNA‐seq reads for each of the samples to the mouse reference genome GRCm38, and Cufflinks to annotate them. The counts of aligned reads to each gene were calculated with HTSeq. In‐house software was used to merge the expression results into a single text file used in the downstream analysis in Matlab (MathWorks). We performed quantile normalization to equalize the data and stabilized them through the log2 transform of the data plus one. The heatmap of the most highly variable transcripts, the hierarchical clustering dendrograms (calculated using the unweighted pair group method with arithmetic mean and Euclidean distance measure), and the PCAs were performed using in‐home functions developed in Matlab (MathWorks).
2025-01-10 | GSE213158 | GEO
Project description:It has been widely accepted that mammalian neural stem cells (NSCs) exist only in the central nervous system. Here, we report that peripheral NSCs (pNSCs) exist outside of the CNS and can be isolated—from mouse embryonic limb, postnatal lung, tail, dorsal root ganglia, and adult lung tissues. Derived pNSCs are distinguished from neural crest stem cells, express multiple NSC-specific markers and exhibit cell morphology, self-renewing capacity, genome-wide transcriptional profile, epigenetic features, and differentiation ability similar to those of control brain NSCs. pNSCs are from Sox1+ cells and originate from neuroepithelial cells. Single-cell RNA sequencing identifies that pNSCs in situ show similar molecular feature to brain NSCs. Furthermore, many pNSCs that migrate out of the neural tube can differentiate into neurons and limited glia cells during embryo development. Our finding of the existence of pNSCs provides previously unidentified insight of the mammalian nervous system development and presents an alternative potential strategy for neural regenerative therapy.
2025-01-10 | GSE213132 | GEO
Project description:Multipotent neural stem cells originating from neuroepithelium exist outside of the mouse central nervous system
| PRJNA879224 | ENA
Project description:Multipotent neural stem cells originating from neuroepithelium exist outside of the mouse central nervous system
| PRJNA879421 | ENA
Project description:Multipotent neural stem cells originating from neuroepithelium exist outside of the mouse central nervous system
| PRJNA636763 | ENA
Project description:Multipotent neural stem cells originating from neuroepithelium exist outside of the mouse central nervous system [scRNA-seq]
| PRJNA879225 | ENA
Project description:Multifocal synchronous or metachronous atypical teratoid rhabdoid tumors (ATRTs) and non-central nervous system malignant rhabdoid tumors (extra-CNS MRTs) are rare cancers. We reviewed the clinical and radiologic characteristics of affected patients seen at our institution. Genotyping and analysis of copy number abnormalities (CNAs) in SMARCB1 were performed in germline and tumor samples. Tumor samples underwent genome-wide DNA methylation and CNA analysis. The median age at diagnosis of 21 patients was 0.6 years. Two-thirds of ATRTs and extra-CNS MRTs were diagnosed synchronously. Although kidney tumors predominated, including two patients with bilateral involvement, at least 30% of cases lacked renal involvement. Histopathologic review confirmed MRTs in all cases and INI1 expression loss in all tumors tested. Fourteen (78%) of 18 patients tested had heterozygous germline SMARCB1 abnormalities. At least one allelic SMARCB1 abnormality was confirmed in 81 and 88% of ATRTs and extra-CNS MRTs, respectively. Unsupervised hierarchical clustering analysis of DNA methylation in 27 tumors and comparison with a reference group of 150 ATRTs classified the CNS tumors (n = 14) as sonic hedgehog (64%), tyrosinase (21%), and MYC (14%). The MYC subgroup accounted for 85% of 13 extra-CNS MRTs. Of 16 paired ATRTs and extra-CNS MRTs, the tumors in seven of eight patients showed a different pattern of genome-wide DNA methylation and/or CNAs suggestive of non-clonal origin. CNS and extra-CNS tumors had an identical SMARCB1 amplification (n = 1) or very similar DNA methylation pattern (n = 1) suggestive of clonal origin. All patients died of tumor progression. The clinical and molecular characteristics of multifocal ATRTs and extra-CNS MRTs are heterogeneous with most patients harboring a cancer predisposition. Although independent tumor origin was confirmed in most cases, metastatic spread was also documented. The recognition of their distinct molecular characteristics is critical in selecting new biologic therapies against these deadly cancers.
| S-EPMC6063764 | biostudies-literature
Project description:Multifocal synchronous or metachronous central nervous system (CNS) and extra-CNS malignant rhabdoid tumors (MRTs) are rare and deadly cancers. To further understand these cancers, we performed genome-wide DNA methylation and CNA analysis in 27 tumors (16 of which were paired samples). 26 samples were newly generated and one sample was from a previously published cohort of MRTs. We compared our results with the previously published reference group of 150 atypical teratoid rhabdoid tumors. We showed that that CNS and extra-CNS MRTs for the most part clustered in distinct methylation groups and had heterogeneous molecular characteristics.
2017-12-12 | GSE107946 | GEO
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. Furthermore, single cell RNA-seq results identify a unique molecular feature of in vivo pNSCs which express NSCs and oligodendroglial markers. Our finding of pNSCs out of CNS expands the field of developmental neuroscience and presents an alternative potential strategy for neural regenerative therapy.
2025-01-10 | GSE151700 | GEO