GSM9498120: Day 1 (D1); Homo sapiens; RNA-Seq
On the day of collection, vascular beds were excised from microfluidic devices with a scalpel, washed 3 times with warm PBS, minced, and immersed in Accutase (Gibco) dissolved with 10 mg/ml Nattokinase (Japan Bio Science Laboratory). The vascular bed compartments of the microfluidic devices were further rinsed with Accutase to dislodge remaining vascular bed fragments. Samples from 4 separate devices were combined per condition and were incubated for 20 min on an orbital shaker at 37°C with mixing by pipetting up and down after 10 minutes. Twice, cell suspensions were passed through 35 µm cell strainers (Falcon) followed by centrifuging and resuspension in fresh EGM-2 media. Samples were processed at the Network Biology Collaborative Centre (NBCC) using the Chromium platform (single cell 3' v3 chemistry; 10X Genomics), followed by sequencing of cDNA libraries on a NovaSeq 6000 (Illumina).