GSM3336595: neuruloid_day7_56CAG; Homo sapiens; RNA-Seq
Two micropatterned glass cover slips with 500μm of diameter neuruloids (BMP4 3ng/ml) were grown in parallel using RUES2 hESCs and the isogenic 56CAG HD line. At day 7, approximately half of each cover slip with neuruloids was scraped off and treated with Accutase (Stemcell technologies) for 10min at 37°C. The remaining neuruloids on the cover slip were fixed for immunofluorescence analysis as quality control. After dissociation the cells were washed three times in PBS+/+ (gibco) with 0.04% BSA and strained through a Flowmi tip strainer (40 μm, cat.no. 136800040). Cell count and viability were determined on a Countess II Automated Cell Counter using trypan blue. The two samples were separately loaded for capture with the Chromium System using the Single Cell 3' v2 reagents (10× Genomics). Following cell capture and lysis, cDNA was synthesized and amplified according to the manufacturer's instructions (10× Genomics). The resulting libraries were sequenced on the Illumina NextSeq500 platform with 150-cycle high output.
GEO Accession
GSM3336595