GSM8222789: ASD WT Stroma 1 (MEH279); Mus musculus; RNA-Seq
Tissues were manually dissociated and then digested for 1 hour at 37°C in 2U/mLCollagenaseB (#11088831001, Roche)/Dispase II (#04942078001, Roche), 50mM HEPES/KOH pH 7.4; 150mM NaCl. Cells were then spun down and resuspended in 2X volume 10%fetal bovine serum(FBS) (#SH30910.03, GE Hyclone) in phosphate buffered saline (PBS) to inactive digestion enzymes, samples were sequentially filtered through 70μm (#22363548, Fisher) and 40μm (#22363547, Fisher) filters to yield single cell suspensions. Data was acquired on a FACS Aria Cellsorter (BD Biosciences) and analyzed using FACSDiva software. DAPIused as a live/dead cell marker. RNA extraction and library preparation was performed according to manufacturer's instructions (single cell 3' v3.1 protocol, 10x Genomics)