title: Illumina HiSeq 2500 sequencing; Impaired Fetal Lung Development can be Rescued by Administration of Extracellular Vesicles Derived from Amniotic Fluid Stem Cells
design: Impaired Fetal Lung Development can be Rescued by Administration of Extracellular Vesicles Derived from Amniotic Fluid Stem Cells
library_construction_protocol: At E14.5, a single cell suspension was obtained from pooled lungs from either control or nitrofen exposed rat fetuses by trypsinization (0.25% Trypsin-EDTA, ThermoFisher, Waltham, MA) for 20 minutes at 37°C humidified chamber. Cells were spun down by centrifugation for 5 minutes at 800g and the pellet was resuspended in DMEM supplemented with 10% FBS and subjected to three serial depletion of fibroblasts by incubation for 1 hour each following an established protocol [Caniggia 1991]. Groups were assigned randomly. Treatment with EVs is with 10% (by volume) Total RNA was extracted using Nucleospin miRNA kit (Macherey-Nagel, Düren, Germany), following supplier recommended protocols. NEBNext small RNA kit was used to construct libraries.