biostudies-arrayexpressDhanya RamachandranHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-14356We aimed to identify specific targets of alternative splicing and gene expression patterns in fetal ECFCs. We performed transcriptome-wide differential splicing and gene expression analyses between cord blood ECFCs from preeclamptic (n=16) and normal pregnancies (n=13).biostudies-arrayexpressNucleic Acid Extraction - Total RNA was purified from ECFC lysates using the RNeasy Plus Mini Kit (Qiagen, Hilden, Germany).Sample Collection - We centrifuged umbilical cord blood and separated plasma for storage at -80°C. After density gradient centrifugation, the cord blood mononuclear cell fraction was plated at 5 x 107 cells/well on collagen coated 6-well plates ((BioCoat; Corning, USA) and incubated at 37 °C and 5 % CO2. Cells were grown with endothelial basal medium (EBM-2, Lonza, Basel, Switzerland) supplemented with 10% fetal bovine serum (FBS, Harvard Bioscience, Harvard Bioscience, Holliston, USA), 1% penicillin/streptomycin (P/S, Bio&Sell, Nürnberg, Germany) and recommended supplements. After 10 to 21 days, ECFC colonies appeared as adherent single layers of cobblestone-shaped cells. ECFC phenotype was confirmed by flow cytometry assessing CD31 (130-117-390; BD Biosciences San Jose, USA), CD45 (555483; BD Biosciences), CD133 (130-090-826; Miltenyi Biotec, Bergisch Gladbach, Germany) and the corresponding isotype controls.Library Construction - rRNA was depleted and PolyA enriched without globin depletion. A strand-specific library was utilized with the protocol NEB Ultra Directional RNA.Sequencing - Samples were sequenced on the NovaSeq PE50 platform. All samples were mapped to human reference genome hg38 and transcriptome Homo_sapiens.GRCh38.79.gtf using STAR aligner version 2.4.2a for mapping. All samples had minimum of 30 million reads after removing counts mapping to rRNA, pseudogenes and Hemoglobin.OrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesData Transformation - Counts (filtered) were transformed into counts per million and expressed as Log2 value. Log2CPM were normalized using TMM (weighted trimmed mean of M−values (to the reference).UnknownTranscriptomicsGenomicsProteomicsIllumina NovaSeq 6000RNA-seq of total RNAHomo sapiensAlternative splicing of CADM1 is associated with endothelial progenitor cell dysfunction in preeclampsiaThilo DörkDhanya RamachandranFrauke von Versen-HöynckRicarda Welz, Dhanya Ramachandran, Bianca Schröder-Heurich, Katja Richter, Robert Geffers, Constantin S. von Kaisenberg, Thilo Dörk, Frauke von Versen-HöynckfalseAlternative splicing and gene expression patterns of fetal cord-blood derived Endothelial Colony Forming Cells (ECFCs) in normal and preeclamptic pregnanciesWe aimed to identify specific targets of alternative splicing and gene expression patterns in fetal ECFCs. We performed transcriptome-wide differential splicing and gene expression analyses between cord blood ECFCs from preeclamptic (n=16) and normal pregnancies (n=13).2025-08-07T00:00:00Z2024-08-12T20:39:50.166Z2024-08-12T20:39:50.166ZE-MTAB-14356ERP163210EFO_0002944EFO_0004170EFO_0009653EFO_0005518EFO_0003816EFO_0004184