GSM9365059: hESC-derived HLO-D21; Homo sapiens; RNA-Seq
On D21, HLOs from three wells of an ultra-low attachment 24-well plate were collected and washed 2x with DPBS (-/-). The HLOs were then dissociated using TrypLE Express for 15 minutes at 37 °C. Following dissociation, the cell suspension was passed through a 70 μm MACS SmartStrainer to obtain a single-cell suspension. Cells were collected in 1% BSA in DMEM, centrifuged at 300 × g for 5 minutes, and resuspended again in 1% BSA in DMEM. The 10× Genomics Chromium equipment and the 3′v3.1 reagent kit were used to create single-cell RNA-Seq libraries in accordance with the manufacturer's instructions. A Qubit fluorometer (ThermoFisher Scientific) and a Tapestation (DNA High sensitivity chip - Agilent Technologies) were used for library quantification and quality evaluation. Libraries were sequenced on an Illumina NovaSeq 6000 for paired-end 28 - 90 reads.