NextSeq 500 paired end sequencing; RNA-seq to investigate the metabolic profile of primary human mesenchymal stromal cells cultured in physiological conditions
RNA-seq to investigate the metabolic profile of primary human mesenchymal stromal cells cultured in physiological conditions
MSCs were seeded at 10,000 cells/cm2 in twelve-well plates in Plasmax. After 24h, media were renewed with fresh Plasmax, supplemented with 10% dialyzed FBS and antibiotics, and cells were incubated at 21% or 1% of O2. After further 48h, cells were harvested for RNA extraction  samples were processed using the QIAshredder/RNeasy Mini Kit (Qiagen) Libraries for cluster generation and DNA sequencing were prepared using Illumina TruSeq RNA Library Preparation Kit. Quality and quantity of the DNA libraries were assessed on an Agilent 2200 TapeStation (D1000 screentape) and Qubit (Thermo Fisher Scientific), respectively.