title: GSM9265254: 1_T1003_Uninfected; Homo sapiens; RNA-Seq
library_name: GSM9265254
library_construction_protocol: Bronchial epithelial cells (BECs) from children were obtained while under general anesthesia for an elective surgery using 4-mm Harrell unsheathed bronchoscope cytology brushes inserted through an endotracheal tube. BECs (passage 2 or 3) were proliferated on transwells under submerged culture conditions using PneumaCult™EX-Plus medium (Stemcell™) for 4-7 days then differentiated to an organotypic pseudostratified ciliated state for 21 days at ALI. BEC cultures were infected on the apical surface with human rhinovirus A16 (RV-A16, Source: ATCC®) at a multiplicity of infection (MOI) of 0.5. Viral suspension was removed from the apical surface after 2 hours of incubation with no subsequent rinsing. RNA was collected from BECs pre-infection and 2 days post-infection for scRNA-seq data. The 10X Genomics Chromium Next GEM Single Cell Fixed RNA Sample Preparation Kit was used to fix the BECs. Samples were stored at -80°C until further post-storage processing and sequencing. Fixed samples were processed according to manufacturer's instructions, using the Chromium Next GEM Single Cell Fixed RNA Sample Preparation Kit (10x Genomics).  Briefly, fixed samples were thawed and hybridized overnight to one of four human probe sets (10x Genomics), with or without additional spiked-in custom rhinovirus probes. Pooled samples were washed and loaded onto the Chromium X (10x Genomics) for partitioning into GEMs.  Gene expression libraries were generated according to the Chromium Fixed RNA Profiling Reagent Kits for Multiplexed Samples user guide.  Sequencing was carried out on a NextSeq 2000 sequencer, using NextSeq 2000 P4 XLEAP-SBS flowcells (Illumina) with a target depth of 10,000 reads/cell.
identifiers.namespace: GEO
identifiers.id: GSM9265254_r1