scRNA-seq of human embryonic stem cell 9 iPSC
Microglia were differentiated to day 45 with minor adaptations to Washer et al (PMID: 36376339) including standardized EB nucleation and use of fibronectin 1mg/ml for attachment at day 6. 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.