Project description:In our study,we examined the effects of naloxone (an opioid receptor antagonist) on NSCs (neural stem cells) proliferation and differenntiation. Briefly, we treated NSCs with naloxone in the proliferation and differentiation system. Naloxone was shown to acclerate NSCs proliferation and increase neural differentiation.To investigate how DNA methylation is affected upon naloxone treatment, cells cultured 3 days with naloxone and cells differentiated with naloxone at D2 were collected, genomic DNA was extracted and subjected to RRBS analysis.
Project description:Primitive neural stem cells (NSCs) could be derived from pluripotent mouse embryonic stem (ES) cells, and then differentiate into definitive-type neural stem cells which resemble NSCs obtained from the central nervous system. Hence, primitive NSCs define an early stage of neural induction and provide a model to understand the mechanism that controls initial neural commitment. In this study, we performed microarray assay to analyze the global transcriptional profiles in mouse ES cell-derived primitive and definitive NSCs and to depict the molecular changes during the multi-staged neural differentiation process. Primitive NSCs derived directly from ESCs in Lif (p-NSC_L), primitive NSCs that were sub-cultured in the presence of Lif and FGF (p-NSC_LF), as well as definitive NSCs derived from primitive NSCs in medium containing FGF and EGF, were collected for RNA extraction and hybridization on Affymetrix microarrays. Mouse ESCs and NSCs obtained from mouse embryonic brain (E11.5) were included for controls. For each cell type, we collected two biological replicate samples for microarray analysis.
Project description:Both Smchd1 deletion and its MommeD43 mutant version cause loss of long-range chromatin interactions in NSCs and a strengthening of more local interactions. We assayed CTCF binding in the relevant genotypes to see if this is related to altered CTCF binding. ChIP-seq using an anti-CTCF antibody to check if chromatin binding of CTCF is altered in Smchd1MD43-GFP/MD43-GFP and Smchd1del/del neural stem cells (NSCs) compared to Smchd1GFP/GFP NSCs.
Project description:Primitive neural stem cells (NSCs) could be derived from pluripotent mouse embryonic stem (ES) cells, and then differentiate into definitive-type neural stem cells which resemble NSCs obtained from the central nervous system. Hence, primitive NSCs define an early stage of neural induction and provide a model to understand the mechanism that controls initial neural commitment. In this study, we performed microarray assay to analyze the global transcriptional profiles in mouse ES cell-derived primitive and definitive NSCs and to depict the molecular changes during the multi-staged neural differentiation process.
Project description:Transcriptional profiling of Human ESCs vs Human iPSCs, Human NSCs-ES vs Human NSCs-iPS iPSCs generated by using different method, and are not very good at Neural differentiation comapred with Human ESCs
Project description:We describe XmaI-RRBS method for rapid and affordable genome-wide DNA methylation analysis, with library preparation taking only four days and sequencing possible within four hours. Small sizes of the XmaI-RRBS libraries allow their multiplexing and sequencing on the benchtop high-throughput machines. Described here is the first RRBS protocol validated for the Ion Torrent Personal Genome Machine. DNA from MCF7 cell line and 6 normal breast samples (total 7 samples) were subjected to reduced representation bisulfite sequencing analysis (XmaI-RRBS) by using Ion Torrent platform.
Project description:Naloxone is an opioid-receptor antagonist used to reverse the effects of opioid.In our study, we treated NSCs (neural stem cells) with naloxone and damgo (a selective μopioid receptor agonist), and examined their effects on NSCs proliferation and differentiation.Naloxone was found to promote NSCs proliferation and neural differentiation, while the effects of damgo was very week. The results suggested that naloxone may work indepedent of opioid receptors. To investigate the underlying mechanisms, RNA-seq was conducted. For proliferation analysis,p8 NSCs were cultured with or without naloxone or damgo for 3 days. For differentiation analysis,p8 NSCs was induced to differentiate with or without naloxone or damgo, samples on D2,4,8 were collected and analyzed.
Project description:Rat NSCs were isolated from developing whole brian of the SD rat embryos at day 13.5. In the NSC/GFP-EMSC co-culture system, the NSCs showed significantly enhanced neuronal differentiation rather than astrocytic commitment, in comparison with the mono-cultured NSCs. In order to reveal the gene expression profiles of NSCs under these two diffferent culture conditions, we used microassrays to examine the global programme of gene expression of NSCs under these two diffferent culture conditions.