Isolation of human ESC-derived cardiac derivatives and embryonic heart cells for population and single-cell RNA-seq analysis

The combination of population and single-cell RNA sequencing analysis using hu-man embryonic stem cell (hESC) differentiation and developmental tissues is a powerful approach to elucidate an organ-specific cellular and molecular atlas in human embryogenesis. This protocol describes (1) cardiac-directed differentia-tion and isolation of hESC-derived cardiac derivatives with fluorescence -acti-vated cell sorting, (2) isolation of human embryonic heart-derived single cardiac cells, and (3) construction of cDNA libraries with Smart-seq2. These allow for the preparation of human developmental samples for comprehensive transcriptional analysis. For complete details on the use and execution of this protocol, please refer to Sahara et al. (2019).

Automated summary

The combination of population and single-cell RNA sequencing analysis in hESC differentiation and developmental tissues is a powerful approach for studying organ-specific cellular and molecular atlas in human embryogenesis. This protocol covers steps including cardiac-directed differentiation, isolation of cardiac derivatives, isolation of single cardiac cells from human embryonic hearts, and construction of cDNA libraries with Smart-seq2 for comprehensive transcriptional analysis of human developmental samples. Refer to Sahara et al. (2019) for detailed information on the use and execution of this protocol.