characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-05-25T00:19:47Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-05-25T00:19:47Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA110462856
characteristics.characteristic: External Id
characteristics.text: Department of Pediatrics and Adolescent Medicine, Medical University of Vienna, Austria
characteristics.characteristic: INSDC center name
characteristics.text: 2023-05-25T00:19:47Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-05-25T00:19:47Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS12560884
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12021:Sample 5
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: HUVEC
characteristics.characteristic: cell_line
characteristics.text: endothelial cell
characteristics.characteristic: cell_type
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: Human endothelial umbilical vein cells (HUVEC), human cardiac microvascular endothelial cells (HCMEC) and the immortalized mesothelial cell line (MeT-5A) were purchased from established vendors (HUVEC and HCMEC from Promocell, Heidelberg, Germany; MeT-5A (ATCC® CRL-9444™) from LGC Standards, Wesel, Germany). Endothelial cells were grown in endothelial cell growth medium (Promocell, Heidelberg, Germany) with supplements and antibiotics according to manufacturer´s instructions. MeT-5A were cultured in Medium 199 (M199, 31150022, Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% (v/v) fetal bovine serum (FBS, Gibco, Thermo Fisher Scientific, Waltham, MA, USA), 1% (v/v) penicillin/streptomycin (P/S, Gibco, Thermo Fisher Scientific, Waltham, MA, USA). Human peritoneal mesothelial cells (HPMC) were isolated from four non-uremic patients and cultured as previously described [Stylianou et al. Kidney Int. 1990], as approved by the Ethic committee of the Medical Faculty, Heidelberg University (S-501/2018). Informed written consent was signed by the patients. The cells were grown in M199 medium supplemented with 10% FBS, 1% penicillin/streptomycin, 0.5 μg/ml insulin, 0.5 μg/ml transferrin, 0.4 μg/ml hydrocortisone and 2 mM L-glutamine (all from Merck, Darmstadt, Germany). Cells were seeded at a density of 2x 105 cells per well on polyester mesh (24 mm Transwell®, 0.4 µm pore size, 6-well type; Corning, MA, USA) under normal culture condition in 3 technical replicates per group. HCMEC and HPMC were pooled (4 donors) before seeding while HUVEC were bought pooled. The insert and outer chamber were filled with 1.5 and 3 mL of cell culture medium, respectively. Cell pellets were collected after reaching plateau. Total RNA isolation was performed using RLT buffer with 1% ß-mercaptoethanol and purified using Micro RNAeasy Kit with on column DNA digestion (Qiagen, Hilden, Germany) following the manufacturer´s procedure. The RNA concentration was measured using Qubit kit (Invitrogen). RNA integrity quantification was performed using RNA screen tape (Agilent, Santa Clara, USA). Detected RNA integrity number (RIN) values of samples were over 9.0 and 500 ng/sample were used for sequencing. The library was prepared using TruSeq Stranded mRNA Library Prep Kit (Illumina, San Diego, USA).
characteristics.characteristic: description
characteristics.text: neonate
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: umbilical vain
characteristics.characteristic: organism part
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: Sample 5
characteristics.characteristic: title