biostudies-arrayexpressFlorian FinkernagelHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-14412Human peritoneal mesothelial cells (HPMCs) were incubated with the conditioned media collected from tumor cells genetically modified to overexpress BCAM.biostudies-arrayexpressSample Treatment - HPMCs are incubated with conditoned media for 48 hrs. Conditioned media: BCAM overexpression was achieved by transient transfection of OVCAR8 cells with BCAM1 vector (Transcript variant 1; OHu20355 – Gencript Biotech; Piscataway, NJ) or BCAM2 vector (Transcript variant 2; OHu07730 – Gencript Biotech, Piscataway, NJ) or empty pCDNA3.1 control (GenScript Biotech, Piscataway, NJ) using TransIT‐X2 (MirusBio/Mobitec; Göttingen, Germany) according to the manufacturer's instructions. OVCAR8 cells were cultured in RPMI 1640 (Cat. #61870044, Thermo Fisher Scientific, Darmstadt, Germany) supplemented with 10% FBS (Cat. #FBS-LE-12A/RES1822, Capricon Scientific, Ebsdorfergrund, Germany). Cells were seeded in 10 cm dishes at a density of 3.5 × 10⁶ cells per dish, and the medium was replaced with serum-free medium after 24 hrs. BCAM-OE cells were incubated for an additional 48 hrs. CM was collected, centrifuged at 300xg for 5 min to remove dead cells, and further clarified by centrifugation at 1500xg for 5 min.Sequencing - Sequencing was performed using Illumina NextSeq 550 v2.5 kits according to manufactures' protocol.Sample Collection - fter 48 hrs, the HPMCs were lysed with RA1 buffer for RNA isolation.Library Construction - “QuantSeq 3′ mRNA-Seq Library Prep Kit FWD for Illumina” (Lexogen, Vienna, Austria)Growth Protocol - In brief, macroscopically tumor-free omentum was digested with trypsin for 30 min, followed by MACS depletion of CD45+ and EpCAM+ cells. Mesothelial cells were cultured in OCMI/5% FCS medium35 for a maximum of 3-5 passages. Human umbilical vein endothelial cells (HUVECs) were cultured using HUVEC Endothelial Growth Media (Cat. #C-22011, PromoCell; St. Louis, USA).Nucleic Acid Extraction - MN NucleoSpin RNA Isolation Kit according to manufacture's instructions.OrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesData Transformation - STAR 2.7.1d onto Ensembl 96. BAM files were anonymized with BAMBoozle.MetabolomicsUnknownTranscriptomicsGenomicsProteomicsNextSeq 550RNA-seq of coding RNAHomo sapiensFlorian FinkernagelSuresh SivakumarRolf MüllerfalseHuman peritoneal mesothelial cell response to conditioned media of BCAM overexpressing OVCAR8 cells (genetic modification)Human peritoneal mesothelial cells (HPMCs) were incubated with the conditioned media collected from tumor cells genetically modified to overexpress BCAM.2025-12-31T00:00:00Z2025-12-31T02:02:16.195Z2024-09-03T17:05:47.694ZE-MTAB-14412E-MTAB-14411EFO_0002944EFO_0004170EFO_0003789EFO_0005518EFO_0003816EFO_0003738EFO_0004184EFO_0003969