SRP413061 PRJNA912190 GSE220972 Investigating the effects of inflammatory cytokines associated with asthma on airway smooth muscle cell contractility Understanding the gene expression changes that are occuring with TGFb1 and IL-13 treatment that result in observed phenotypic changes in bronchial smooth muscle shortening/remodeling. Overall design: Normal human bronchial smooth muscle cells (hBSMCs) were obtained from Lonza (CC-2576) and cultured using SmGMTM-2 BulletKitTM (Lonza CC-3182). To seed the microtissues: Cells were then suspended in a 2%1 mg/mL neutralized type 1 collagen solution then dispensed at a density of 50,000 cells per well into the plate over ice. Cantilever arrays were immediately added to the well ensuring the ends of each cantilever were fully submerged and incubated at 37? for 15 min to allow for gelation. Growth media was then added to the seeded wells and plates were incubated overnight to allow for initial tissue formation (Day 0). Following 24h of incubation (Day 1) cantilever arrays were transferred into a fresh 96 well plate containing serum free mediahBSMC growth media with reduced serum (0.5% FBS) and the indicated recombinant cytokines at 10 ng/mL (see Supplementary Table X for catalog numbers). Microtissues were incubated for 3 days after which the media and cytokines were replenished (Day 4). After an additional three days of treatment (Day 7), microtissues were used for desired endpoint assessment, which includes brightfield imaging for contractility differences and lysing tissues in Qiagen Buffer RLT for RNA isolation, performed using the Qiagen RNEasy kit. GSE220972 pubmed 37206658