title: GSM5420297: MSCs - 6-OHDA treated 1; Mus musculus; RNA-Seq
library_construction_protocol: For the isolation of adipose tissue cells, tissue was collected into PBS, cut into small pieces and incubated with Liberase TM (2.5µg/ml, Roche) and DNase I (20U/ml; Roche) for 1h at 37°C under gentle agitation. A single cell suspension was obtained by passage through a 100μm cell strainer (Thermo Fisher Scientific) and centrifugation was used to separate the stromal vascular fraction from the adipocyte fraction. Erythrocytes were lysed with red blood cell lysis buffer (eBioscience) and removed by centrifugation. Cell suspensions were stained and populations were gated on live cells and defined as MSCs: CD45- Sca-1+ CD31- Pdgfra+. Cells were sorted using FACSFusion (BD Biosciences). Total RNA from sorted cells was extracted using RNeasy micro kit or RNeasy mini kit (Qiagen) according to the manufacturer's protocol. RNA libraries were prepared for sequencing using standard Illumina protocols