GSM5839832: WTC iBMEC 2D 2 mL 3 [A35]; Homo sapiens; RNA-Seq
3D experiments: iBMECs subcultured for one hour were detached using Accutase and then seeded into 150 μm diameter channels patterned in 7 mg/mL genipin-crosslinked type I collagen (Corning) as previously reported (PMID: 30391800). 3D microvessels comprised of BC1 iBMECs were cultured for two days, and then treated with and without 5 ng/mL human recombinant TNF-alpha (R&D Systems) for 24 hours. RNA was collected at the following time points: three days after passing for WTC iPSCs, two days after subculture or seeding into 3D microvessels for WTC iBMECs in 2D and 3D, two days after passing following differentiation for WTC iPCs and iECs, three days after subculture or seeding into 3D microvessels for BC1 iBMECs in 2D and 3D, and two days after passing for HDMECs. Cells were washed with 1xPBS and lysed using RLT buffer supplemented with β-mercaptoethanol, before isolating RNA using a RNeasy Mini Kit (Qiagen) with DNase I digestion. RNA libraries were prepared for sequencing using standard Illumina protocols conducted by Novogene.