GW17 oRG after FCAS
After FACS, outer radial glial cells in single-cell suspension were either directly subjected to single-cell RNA sequencing or plated onto matrigel coated plates and allowed for spontaneous differentiation with or without LIF treatment. After 28 days of cultures, cells were resuspended by papain and subjected to single-cell RNA sequencing. Single-cell capture from live cells was performed following manufacturer's instructions from 10 Genomics for the Chromium Next GEM Single Cell 3' Kit v3.1. Libraries were sequenced as per manufacturer recommendation on Illumina NovaSeq 6000 sequencer, targeting 30,000 reads per cell.