biostudies-arrayexpressNora Doleschallsynthetic constructAgilent Feature ExtractionNonehttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-14672This experiment was used to assess levels of intra- and extracellular/secreted miRNAs in HUVEC cell cultures, and to additionally observe the variability in miRNA expression landscape between various cell lines and culture conditions.biostudies-arrayexpressSample Collection - Conditioned media was removed from cell cultures, and cleared of debris by centrifugation at 5000g. Cells were then washed with HBSS buffer then TRI reagent was added to each well. Cell lysates and conditioned media were stored at -80 until RNA extraction.Hybridization - Hybridisation was performed over 20 hours at 55 degrees C, rotating 20rpm in Agilent hybridisation ovenLabeling - Cy3-pCp labelling was performed according to Agilent recommendationsScaning - Agilent Microarray Scanner (G2505C) was used for array scanning and Agilent Feature Extraction software used for image processing.Growth Protocol - Cells were grown for 6 hours in low-serum conditions in four different types of medium. MCDB, its reduced growth factor containing variant, MCDB-448 S (1% FBS, no recombinant EGF, bFGF, nor ITS), HIM-V medium (AIM-V from 449 ThermoFisher completed with 1% filtrated, heat inactivated FBS, 1 ng/mL bFGF, 2 ng/mL 450 EGF and 7.5 U/mL heparin), and EGM-2MV (Lonza, CA, USA) were used.Nucleic Acid Extraction - smallRNA enriched RNA extraction was performed using Qiagen miRNeasy kitMIAME ScoreRaw DataOrganizationAssays and DataProcessed DataMAGE-TAB FilesArray DesignsData Transformation - Agilent Feature Extraction software was used to assign probes to genes and to perform background correction and error evaluation for individual probesUnknownTranscriptomicsGenomicsProteomicsAgilent Microarray Scanner G2505CPCAgilent Hybridisation Oven G2545ANonemicroRNA profiling by arraysynthetic constructLászló CervenakNora DoleschallfalsemiRNA microarray of 4 primary HUVEC lines and their conditioned media cultured in 4 different cell culture mediaThis experiment was used to assess levels of intra- and extracellular/secreted miRNAs in HUVEC cell cultures, and to additionally observe the variability in miRNA expression landscape between various cell lines and culture conditions.2025-06-30T00:00:00Z2024-12-09T18:27:11.926Z2024-12-09T18:27:11.926ZE-MTAB-14672EFO_0002944EFO_0003814EFO_0000753EFO_0003813EFO_0003789EFO_0005518EFO_0003816EFO_0003815