Project description:<p>During development of the human brain, multiple cell types with diverse regional identities are generated. Here we report a system to generate early human brain forebrain and mid/hindbrain cell types from human embryonic stem cells (hESCs), and infer and experimentally confirm a lineage tree for the generation of these types based on single-cell RNA-Seq analysis. We engineered <i>SOX2<sup>Cit/+</sup></i> and <i>DCX<sup>Cit/Y</sup></i> hESC lines to target progenitors and neurons throughout neural differentiation for single-cell transcriptomic profiling, then identified discrete cell types consisting of both rostral (cortical) and caudal (mid/hindbrain) identities. Direct comparison of the cell types were made to primary tissues using gene expression atlases and fetal human brain single-cell gene expression data, and this established that the cell types resembled early human brain cell types, including preplate cells. From the single-cell transcriptomic data a Bayesian algorithm generated a unified lineage tree, and predicted novel regulatory transcription factors. The lineage tree highlighted a prominent bifurcation between cortical and mid/hindbrain cell types, confirmed by clonal analysis experiments. We demonstrated that cell types from either branch could preferentially be generated by manipulation of the canonical Wnt/beta-catenin pathway. In summary, we present an experimentally validated lineage tree that encompasses multiple brain regions, and our work sheds light on the molecular regulation of region-specific neural lineages during human brain development.</p>
Project description:Global gene expression profiling was performed using RNA from human embryonic neural stem cells (hENSCs) and adult human olfactory bulb-derived neural stem cells (OBNSCs) to define a gene expression pattern and signaling pathways that are specific for each cell lineage. Subtractive gene expression profiling between both cell lineages provides a list of potential genes that are related to their multipotentiality, proliferation, migration, and alternative signaling pathways. To confirm the validity of our DNA microarray data, our results were compared with data from various databases. The gene expression profile of adult olfactory bulb neural stem cells (n=6) was compared with that of human embryonic neural stem cells (n=3).
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: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 primary Neural Plate Border Stem Cells (pNBSCs) isolated from neural folds of E8.5 mouse embryos. Here we provide single cell RNA-sequencing data of neural tissue derived from two E8.5 mouse embryos. After manual isolation and enzymatic separation E8.5 neural tissue was single cell sorted and RNA sequencing was performed following the Smart-seq2 protocol. In sum, cultured pNBSCs and E8.5 neural tube cells share a similar regional identity and expression signature suggesting that pNBSCs might correspond to an endogenous progenitor in this area of the developing brain.
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 array-based expression data of (i)NBSCs and CNS- and neural crest progeny derived thereof. The former comprise radial glia-type stem cells, while the latter are neural crest and mesenchymal stem cell-like cells. The data provided reveal that (i)NBSCs can be directed into defined neural lineages and that iNBSCs pass through successive developmental stages. These data support the notion that it is possible to reprogram human adult cells into expandable, multipotent NBSCs that define a novel embryonic neural stem cell population in human and mouse.