GSM9125075: Cortex, KO, D43, rep1; Homo sapiens; RNA-Seq
Cortical organoids were dissociated using the papain dissociation system per the manufacturer's instructions. In order to increase cell viability, solutions were oxygenated with 95% O2:5% CO2. In brief, HBSS was used to wash the organoids, then they were dissected into small pieces in the oxygenated papain solution. This solution was then re-equilibrated with 95% O2:5% CO2 and placed in a 37 water bath for an hour. The solution was lightly shaken every 20 minutes during the hour incubation period. After the incubation period was over, light trituration was performed to produce a single cell suspension and albumin-ovomucoid inhibitor solution was used to inactivate the papain. hCOs were then subjected to the 10X Genomics Chromium platform. To analyze the transcriptomic profile changes induced by KIAA0319 knockout, scRNA-seq was performed using the D43 hCOs samples with or without knockout of KIAA0319 gene (n = 5 WT and n = 5 KIAA0319 -/- hCOs from one batch). Single-cell suspensions were generated and processed using the 10x Genomics Chromium Single Cell 3' Reagent Kit (v3.1) according to the manufacturer's instructions. Libraries were constructed using 10x Genomics Chromium technology targeting the 3′ end of transcripts.