Project description:We describe a so far uncharacterized, embryonic and self-renewing Neural Plate Border Stem Cell (NBSC) population with the capacity to differentiate into central nervous and neural crest lineages. NBSCs can be obtained by neural transcription factor-mediated reprogramming (BRN2, SOX2, KLF4, and ZIC3) of human adult dermal fibroblasts and peripheral blood cells (induced Neural Plate Border Stem Cells, iNBSCs) or by directed differentiation from human induced pluripotent stem cells (NBSCs). Moreover, human (i)NBSCs share molecular and functional features with an endogenous NBSC population isolated from neural folds of E8.5 mouse embryos. Upon differentiation, iNBSCs give rise to either (1) radial glia-type stem cells, dopaminergic and serotonergic neurons, motoneurons, astrocytes, and oligodendrocytes or (2) cells from the neural crest lineage. Here we provide single cell RNA-sequencing data of two primary mouse Neural Plate Border Stem Cell Lines (pNBSCs). pNBSCs were single cell sorted and RNA sequencing was performed following the Smart-seq2 protocol. In sum, pNBSCs and iNBSCs share a similar regional identity, expression signature and analogous differentiation dynamics on the single-cell-level, suggesting the presence of a transient, NBSC-like progenitor during the neurulation stage of mouse and likely also human embryos.
Project description:Sall4 is a stem cell factor which is important for embryogenesis. We have genetically modified Sall4 in mouse embryonic stem cells to access the transcriptional changes. There are three different genetic modifications for the ES cells in the form of Sall4 Knockout (KO), Sall4 Zinc Finger Cluster 4 Mutation (ZFC4mut) and Sall4 Zinc Finger Cluster 1-2 Deletion (ZFC1-2Δ) respectively that we have considered for our study. Cells were subjected to neural differentiation and directly lysed on the plate at the appropriate timepoint and RNA was sequenced.
Project description:We have examined the nuclear (nuc) and cytoplasmic (cyt) polyA+ transcriptomes of undifferentiated mouse embryonic stem cells (un) and cells differentiated to neural precursors (d5) using strand-specific RNA-Seq. The 46C mouse embryonic stem cell line was used for this study. Two cell types were examined: undifferentiated mouse embryonic stem cells (un) and cells differentiated to neural precursors (d5). For each cell type, cells were fractionated to nuclear and cytoplasmic components. RNAs were extracted from each component and were fragmented enzymatically for library construction. For each cell type and component, strand-specific RNA-Seq libraries were generated using at least two different fragmentation protocols.
Project description:Neural stem cells from different brain regions show differencies in gene expression patterns and physiological functions. We used microarrays to find differential gene expression between the neuralstem cells from the subventricular zone of lateral ventricle and the subventricular zone of third ventricle. Cultured neural stem cells from embryonic day 17.5 mouse embryos were selected for RNA extraction and hybridization on Affymetrix microarrays. We sought to obtain embryonic neural stem cell cultures from three indipendent females.