GSM2071702: 34-NPS1; Homo sapiens; RNA-Seq
Human fetal kidneys were collected from elective abortions. Fetal gestational age ranged from 15 to 19 weeks. All studies were approved by the local ethical committee and informed consent was given by the legal guardians of the patients involved according to the declaration of Helsinki. Each tissue was processed for immunohistochemical (IHC) staining and formation of single cell suspension. Single cells suspensions were suspended in Iscove’s Modified Dulbecco’s Medium (IMDM) medium supplemented with 10% Fetal Bovine Serum (FBS) and growth factors: 50 ng/ml of basic Fibroblast Growth Factor (bFGF), 50 ng/ml of Epidermal Growth Factor (EGF) and 5 ng/ml of Stem Cell Factor (SCF) (R&D Systems) and plated in flasks. Cells were incubated used for FACS sorting of cell subpopulations. Bulk total RNA was prepared from ~1.5*10^5 cells using the Direct-zol RNA MiniPrep kit (Zymo Research) and stored in -80C. RNA was quantified on an Agilent BioAnalyzer (Agilent Technologies) and aliquots of 270-500ng were made into cDNA libraries using the TruSeq mRNA-Seq library kit (Illumina). RNA libraries were prepared for sequencing using standard Illumina protocols. TruSeq RNA-Seq.
GEO Accession
GSM2071702