GSM8539229: human cerebellar organoids, cell line: 1208-2; Homo sapiens; RNA-Seq
Three organoids from each condition were pooled at indicated day, incubated in 30 U/mL papain enzyme solution (Worthington Biochemical, LS003126) and 0.4% DNase (12,500 U/mL, Worthington Biochemical, LS2007) at 37 °C for 45 minutes. Organoids were triturated with a P1000 pipette and were resus-pended in FBS followed by trituration with a P200 pipette tip. Cells were resuspended in PBS with 0.4% DNase and filtered through a 70 μm Flowmi Cell Strainer (Bel-Art, H13680-0070). The samples were centrifuged at 200 x g for 3 minutes and were resuspended in Parse Cell Buffer containing BSA. Cells were then fixed using the Parse Biosciences Cell Fixation (Parse Biosci-ences, ECF2001) kit and stored at −80°C. To target 7,000 cells after recovery, approximately 11,600 cells were loaded per lane on a Chromium Single Cell 3′chip (Chromium Next GEM Chip G Single Cell Kit, 10x Genomics, PN-1000127) and cDNA libraries were generated with the Chromium Next GEM Single Cell 3ʹ GEM, Library & Gel Bead Kit v3.1 (10x Genomics, PN-1000128), according to the manufacturer's instructions. The library was sequenced using the Illumina NovaSeq S4 2 × 150 bp at Admera Health