scRNA-seq of human neural brain organoids patterning reproducibility screening
scRNA-seq of human neural brain organoids patterning reproducibility screening
After d21 in culture 3 organoids per condition per cell line were dissociated individually using CyBio FeliX liquid handler robot with thermoshaker. For dissociation we used a papain-based dissociation kit (Miltenyi Neural dissociation kit). Each organoid was dissociated using 410uL of enzyme mix 1 and 6uL of enzyme mix 2. Then single cell suspension from the same condition were pooled together (H1 and H9; WTC and WIBJ2).  Each individual single cell suspension has been followed by  fixation and  permeabilization procedures, which were performed according to the manufacturer specification (ParseBiosciences, cell fixation kit v3, CF100).  Collected samples were processed for highly multiplexed single-cell RNA sequencing (scRNA-seq) using a split-pool combinatorial barcoding kit (ParseBiosciences, WT Mega kit v3, MG100).  