scRNA-seq of human embryonic stem cell 9 iPSC
Midbrain astrocytes were differentiated from day33 midbrain floorplate progenitors above with minor adaptations to Perriot et al., and harvested at day 125. ESC were captured from freshly dissociated colonies, while all differentiated cryopreserved cells were thawed, depleted for dying cells with AnnexinV microbeads (Mitlenyii, Dead cell removal kit), and debris was floated with a density cushion (4% BSA in L15, adjusted to pH7). Cells were hashtagged, pooled, and captured on 10x Chromium 3 Gene Expression ~20K paired end reads/cell following standard methods.