GSM5885681: human induced pluripotent stem cell-derived 3D heart organoids D10; Homo sapiens; RNA-Seq
Epicardioids were dissociated with papain as described in methods of Meier et al. (2021), filtered through 40 μm filter, washed with 5 mL 1% BSA (Gibco; 15260037) in PBS-/-, centrifuged for 3 min at 200 g, and resuspended in 0.04% BSA in PBS for counting. After counting, 8 000 cells for each sample (targeted cell recovery) were processed using Chromium Single Cell 3' GEM, Library & Gel Bead Kit v3 (10x Genomics; PN-1000092), the Chromium Chip B Single Cell Kit (10x Genomics; PN- 1000073), and the Chromium i7 Multiplex Kit v2 (10x Genomics; PN-120262) for samples d2-d15 and Chromium Next GEM Single Cell 3' Library & Gel Bead Kit v3.1 (1000128, 10x Genomics) Chromium Single Cell G Chip Kit (1000127, 10x Genomics) and Single Index Kit T Set A (PN-1000213, 10x Genomics) for sample d30 to generate Gel Bead-In-EMulsions (GEMs) and single cell sequencing libraries. 8 000 cells for each sample (targeted cell recovery) were processed using Chromium Single Cell 3' GEM, Library & Gel Bead Kit v3 (10x Genomics; PN-1000092), the Chromium Chip B Single Cell Kit (10x Genomics; PN- 1000073), and the Chromium i7 Multiplex Kit v2 (10x Genomics; PN-120262) for samples d2-d15 and Chromium Next GEM Single Cell 3' Library & Gel Bead Kit v3.1 (1000128, 10x Genomics) Chromium Single Cell G Chip Kit (1000127, 10x Genomics) and Single Index Kit T Set A (PN-1000213, 10x Genomics) for sample d30 to generate Gel Bead-In-EMulsions (GEMs) and single cell sequencing libraries. Libraries were pooled and sequenced using the NovaSeq (Ilumina; S1 flow cell) with 150bp paired end reads with 28 cycles for read1, 91 cycles for read2 8 cycles for i7 and 0 cycles for i5, and with a read depth of at least 25-30 000 pair reads per cell.