{"database":"biostudies-arrayexpress","file_versions":[],"scores":null,"additional":{"omics_type":["Metabolomics","Unknown","Transcriptomics","Genomics","Proteomics"],"submitter":["Sarah Boutom"],"instrument_platform":["Illumina NovaSeq 6000","Qubit 4 Fluorometer"],"study_type":["RNA-seq of coding RNA"],"organism":["Homo sapiens"],"species":["Homo sapiens"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/E-MTAB-15954"],"description":["Mechanisms guiding the induction of blood-brain barrier (BBB) properties in central nervous system (CNS) endothelial cells during human development are incompletely understood. To explore induction of reduced vesicular endocytosis and transcytosis properties in a human in vitro model of the BBB, we used human pluripotent stem cell (hPSC)-derived endothelial progenitor cells (EPCs) in which Wnt/β-catenin signaling was activated to generate hPSC-derived CNS-like ECs (hPSC-CECs). We assessed the effects of Notch signaling through overexpression of the Notch1 receptor intracellular domain (N1ICD). N1ICD overexpression in hPSC-CECs was induced by treatment with doxycycline (Dox), and hPSC-CECs were sorted into subpopulations by FACS based on high or low surface expression of CD144/VE-cadherin (CD144hi or CD144lo, respectively). Several negative control replicates treated with PBS (nearly all of which were CD144hi), were included as well. Our findings indicate that Notch signaling differentially regulates vesicular endocytosis and transcytosis-related genes in a human model of the developing BBB, contributing to our understanding of how Notch signaling induces these specific BBB properties in this model of human CNS EC development."],"repository":["biostudies-arrayexpress"],"sample_protocol":["Library Construction - Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification.","Nucleic Acid Extraction - Cells were lysed with TRIzol reagent (Invitrogen) for extraction of total RNA, which was subsequently purified using the Direct-zol RNA Miniprep Kit (Zymo Research). Samples were incubated with RNase-free DNase I (Qiagen) to eliminate residual gDNA during RNA Miniprep protocol. RNA was eluted with RNase-free water and concentration was determined using the Qubit 4 Fluorometer (Invitrogen) and Qubit RNA High Sensitivity Assay Kit (Invitrogen). Purified RNA samples (≥200 ng) were sent to Novogene Corporation (Sacramento, CA) for library preparation and mRNA sequencing.","Sample Collection - hPSC-CECs treated with doxycycline (Dox) or PBS were sorted into subpopulations by FACS based on level of surface CD144 expression (CD144hi and CD144lo).","Sequencing - Illumina NovaSeq 6000 was used for RNA sequencing."],"figure_sub":["Organization","MINSEQE Score","Assays and Data","MAGE-TAB Files"],"pubmed_authors":["Sarah Boutom"],"additional_accession":[]},"is_claimable":false,"name":"Notch signaling activation in hPSC-derived CNS-like endothelial cells induces differential expression of blood-brain barrier-related genes","description":"Mechanisms guiding the induction of blood-brain barrier (BBB) properties in central nervous system (CNS) endothelial cells during human development are incompletely understood. To explore induction of reduced vesicular endocytosis and transcytosis properties in a human in vitro model of the BBB, we used human pluripotent stem cell (hPSC)-derived endothelial progenitor cells (EPCs) in which Wnt/β-catenin signaling was activated to generate hPSC-derived CNS-like ECs (hPSC-CECs). We assessed the effects of Notch signaling through overexpression of the Notch1 receptor intracellular domain (N1ICD). N1ICD overexpression in hPSC-CECs was induced by treatment with doxycycline (Dox), and hPSC-CECs were sorted into subpopulations by FACS based on high or low surface expression of CD144/VE-cadherin (CD144hi or CD144lo, respectively). Several negative control replicates treated with PBS (nearly all of which were CD144hi), were included as well. Our findings indicate that Notch signaling differentially regulates vesicular endocytosis and transcytosis-related genes in a human model of the developing BBB, contributing to our understanding of how Notch signaling induces these specific BBB properties in this model of human CNS EC development.","dates":{"release":"2026-02-17T00:00:00Z","modification":"2026-02-23T15:12:34.031Z","creation":"2025-11-01T17:15:30.21Z"},"accession":"E-MTAB-15954","cross_references":{"ENA":["ERP183498"],"EFO":["EFO_0002944","EFO_0004170","EFO_0005518","EFO_0003738","EFO_0004184"]}}