Abstract 16184: Single Cell RNA Sequencing Reveals Dynamic and Heterogeneous Changes of Transcriptome During Cardiac Differentiation in vitro
Human induced pluripotent stem cells-derived cardiomyocytes (iPSC-CMs) are expected to be clinically applicable, especially in the field of disease modeling and drug screening. However, one of the major problems to be overcome is their immaturity and heterogeneity, because these problems hinder the acquisition of stable experimental results for the clinical use. To overcome these problems, we performed the single cell RNA sequencing during cardiac differentiation in vitro and explored deeply the insights underlining cardiac differentiation. We successfully defined the differentiation state of cells at single cell level by principle component analysis (PCA) and largely recapitulate the cardiac development process with dynamic changes of transcriptome during the differentiation. In addition, by combination with weighted gene co-expression network analysis (WGCNA), we found out that known core transcriptional factors, such as TBX5, GATA4, MEF2C, and Nkx2.5, were co-regulated in the primitive CMs, but were differently regulated in the late differentiation stage, implying that a novel mechanism regulated the cardiac maturation processes. In addition, PCA during cardiac differentiation clarified the heterogeneous development of iPSC-CMs and we identified a novel surface marker, by which we could enrich well-matured CMs from heterogeneous population of differentiated CMs. Taken together, these results suggested that high resolution analyses by single cell RNA sequencing could be applicable not only to deeply understand the cardiac differentiation processes, but also to discover a novel important gene regulation for understanding the immaturity and heterogeneity during differentiation in vitro.
Author Disclosures: S. funakoshi: None. M. Nomura: None. C. Okubo: None. K. Miki: None. T. Kimura: None. S. Yamanaka: Consultant/Advisory Board; Modest; iPierian, iPS Academia Japan, Megakaryon Corporation, and Healios K.K.. A. Watanabe: None. Y. Yoshida: None.
- © 2014 by American Heart Association, Inc.