GSM8885065: Cardiac Organoid, D6_600um; Homo sapiens; RNA-Seq
Cardioids were isolated by first scraping, aspirating, and discarding residual cells and tissue within the wells. Only beating cardioids (~ 15 – 25 cardioids) were collected from each well of 6-well plates. For timepoints prior to Day 6, samples were dissociated with Accutase and quenched with RPMI/B27 minus insulin medium, while samples from Day 6 onward were dissociated using cardiomyocyte dissociation medium (Stem Cell Technologies, cat. no. 05025) and quenched with RPMI/B27 complete medium. Cells were singularized by repeated pipetting and passing through a cell strainer. Single-cell RNA sequencing libraries were prepared using the Chromium Single Cell 3′ reagent kit v3 (10x Genomics, PN-1000075) according to the manufacturer's protocol. Cells were diluted into the Chromium Single Cell A Chip to obtain approximately 6,000 single-cell transcriptomes. After preparation, libraries were sequenced on a NextSeq 500 (Illumina) with 75-cycle high-output kits (Index 1 = 8, Read 1 = 26, and Read 2 = 58).