GSM5034379: SEAM-Control; Homo sapiens; RNA-Seq
differentiated SEAM cultures were dissociated in the incubator for 1-hour at 37 °C, 20% O2, 5% CO2 using 2.5 mg/ml Collagenase 2 (Worthington Biochemical Corporation) in Hanks' Balanced Salt Solution with 3mg/ml DNase I Solution (STEMCELL Technologies) and 2uM Thiazovivin ROCK Inhibitor. The harvested cells were washed with Dulbecco's Modified Eagle Medium: Nutrient Mix F-12 (DMEM/F12, Life Technologies), containing 0.1% BSA, 3mg/ml DNase I Solution and 2 uM Thiazovivin and then filtered with a 40um cell strainer (Corning). Cells were frozen down prior to single-cell RNA sequencing in 90% Heat Inactivated Fetal Bovine Serum (FBS, Sigma) and 10% DMSO (COMPANY) using Mr. Frosty containers. Chromium Single Cell 3' Library and Gel Bead Kit v3.0 using an input of ~10,000 cells, The samples were sequenced on the Novaseq 6000 Illumina sequencer with S4 flow cell, (100/paired end reads) targeting a depth of 50,000-100,000 reads per cell using v3 chemistry 10X Genomics Chromium controller v3.16 alined to GRCh38.100 and SARS-CoV-2 USA-WA1
GEO Accession
GSM5034379