GSM4681839: ARPKD2; Homo sapiens; RNA-Seq
The HO cultures were placed in a 1 ml solution containing 1 mg/ml collagenase and 1 mg/mg dispase (Worthington-Biochem, Lakewood, NJ), and then incubated for 15 to 30 minutes at 37°C with shaking. The cells were further dispersed by gentle pipetting with a fire-polished glass pipette, and then filtered through a 40 μm cell strainer (PluriSelect, San Diego, CA). The single cell suspension was visually inspected under a microscope, cells were counted using a Scepter™ 2.0 Handheld Automated Cell Counter (EMD çMillipore, Burlington, MA), and then resuspended in PBS with 0.01% BSA. Single cells were captured in droplets with barcoded beads using the 10x Genomics Chromium system (Pleasanton, CA). For comparison at  primary HO stage, Control and ARPKD Cellular suspensions (1x106/ml) were then loaded onto the  twon 10× Chromium instrument (10× Genomics); and were then sequenced according to the manufacturer's instructions. 3 ARPKD and 3 control organoid samples were separately sequenced on two lanes of an Illumina HiSeq2500 Rapid Mode instrument, yielding 889M total reads (Contrl=C24 and ARPKD=P42). For compare differentiation trajectory between control and ARPKD, cells from 6 groups sampels including: iPSC (C1, C2 and C3), HB (C1, C2 and C3), HOs (C1, C2 and C3), iPSC (M1, M2 and M3), HB (M1, M2 and M3), HOs (M1, M2 and M3), were barcoded by total 6 cell hashing antibodies according to the manufacturer's protocol (BioLegend TotalSeq™). Then, the pooled cells were captured using two 10x Single Cell 3' Reagent Kit v3 runs; and sequenced on two lanes of an Illumina HiSeq2500 instrument, which yielded 660 M total reads that covered 10,000 cells in total.
GEO Accession
GSM4681839