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GEO help: Mouse over screen elements for information. |
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| Status |
Public on Jun 02, 2023 |
| Title |
Prospective identification and characterization of multipotent peripheral neural stem cells out of mammalian central nervous system |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
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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| Overall design |
12 samples were analyzed
elpNSCs-early, mouse early embryonic limb (el) peripheral Neural Stem Cell (pNSC), 2 replicates
alpNSCs-early, mouse early adult lung (al) peripheral Neural Stem Cell (pNSC), 2 replicates
elpNSCs-late, mouse late embryonic limb (el) peripheral Neural Stem Cell (pNSC), 2 replicates
alpNSCs-late, mouse late adult lung (al) peripheral Neural Stem Cell (pNSC), 2 replicates
Control-NSC, mouse control Neural Stem Cell (NSC), 2 replicates
Fibroblasts, mouse fibroblast, 2 replicates
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| Contributor(s) |
Han D, Jeong H, Hemmer K, Tsytsyura Y, Wu G, Kim K, Renner H, Han DW, Gerovska D, Araúzo-Bravo MJ, Klingauf J, Schwamborn JC, Adams RH, Schöler HR |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
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| Submission date |
Jun 02, 2020 |
| Last update date |
Jun 03, 2023 |
| Contact name |
Marcos J. Araúzo-Bravo |
| E-mail(s) |
mararabra@yahoo.co.uk
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| Phone |
+34 943 00 6108
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| Organization name |
Max Planck Institute for Molecular Biomedicine
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| Department |
Cell and Developmental Biology
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| Lab |
Computational Biology and Bionformatics
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| Street address |
Rogentstrasse
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| City |
Muenster |
| ZIP/Postal code |
48149 |
| Country |
Germany |
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| Platforms (1) |
| GPL6885 |
Illumina MouseRef-8 v2.0 expression beadchip |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA636763 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE151649_MatrixNon-Normalized.txt.gz |
1.3 Mb |
(ftp)(http) |
TXT |
| GSE151649_RAW.tar |
3.1 Mb |
(http)(custom) |
TAR |
| Processed data included within Sample table |
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