GSM8794316: iNs and iMGLs cultured with CLU WT iAs; Homo sapiens; RNA-Seq
Cells were dissociated with a 1:1 mixture of warm trypsin-EDTA:cold StemPro Accutase Cell Dissociation Reagent for 10min at 37°C. Trypsin was quenched with Trypsin Neutralization Solution (ScienCell, 0113-2) prior to collection of cell suspension. Four independent culture wells were combined for each culture condition. Cell suspensions were centrifuged for 5 min at 300 x g and resuspended in 1 mL of BrainPhys media. The cell pellet was triturated gently with a p1000 tip and passed through a 40µM Flowmi Cell Strainer (Millipore, BAH136800040-50) to remove debris. Cell suspensions were centrifuged at 200 rcf for 5min and resuspended in 0.04% BSA in PBS. Washes were repeated two more times. At the end of the final wash, cell suspension was passed through another 40µM Flowmi Cell Strainer. Each triple-culture was loaded onto an individual 10X Chromium Chip well (60,000 cells per well) and resulting emulsions were used to generate scRNAseq libraries using the Chromium Next GEM Single Cell 3' v3.1 chemistry. Libraries were multiplexed and sequenced on a NovaSeq to an average depth of >20K reads per filtered barcode before filtering.