GSM5828137: CHDCases; Homo sapiens; RNA-Seq
The cell lines were differentiated to hiPS-CMs as previously described (Helle et al., 2021). Briefly, Wnt-pathway activation and inactivation were used to induce differentiation, and metabolic selection with media containing lactate instead of glucose was used to ensure purity of the CM culture. The cells were processed for single-cell sequencing as described previously (Helle et al., 2021). The cells were detached using Multi Tissue Dissociation Kit 3 (130-110-204, Miltenyi Biotec), according to the manufacturer's instructions. The cells were washed once with PBS containing 0.04% BSA, and then resuspended in PBS with 0.04% BSA to a concentration of 2.0 × 10^6 cells/ml. The cells were passed through a 35 μm strainer (352235, Corning) and combined into two samples (one containing cells from healthy individuals, one containing patient cells) and placed on ice until continuation of the 10× Genomics Single Cell Protocol. The Chromium Single Cell 3′ RNAseq run and library preparation were made using the Chromium Single Cell 3′ Reagent version 3 chemistry.