Reference of endogenous and pluripotent stem-cell derived cardiomyocyte maturation

The purpose of this study was to understand differences between maturation processes in endogenous and pluripotent stem cell-derived cardiomyocytes. To this end, we first expanded our previous reference of endogenous cardiomyocyte maturation, isolated by LP-FACS (see Kannan et al., bioRxiv 2020, and GSE147807). We subsequently generated PSCs from the same background strain and differentiated to CMs. CMs were isolated by conventional FACs. Overall design: We isolated CMs from mice at the following timepoints (using LP-FACS for all timepoints from p8 onwards): e14, e18, p0, p1, p4, p8, p11, p14, p15, p18, p22, p28, p35, p56, and p84. We isolated PSC-CMs at D8, D10, D12, D15, D18, D25, D30, and D45.

本研究旨在探究内源性心肌细胞与多能干细胞诱导分化心肌细胞的成熟进程差异。为此，我们首先拓展了此前建立的内源性心肌细胞成熟相关参考数据集——该数据集通过LP-FACS分选分离得到（详见Kannan等2020年发表于bioRxiv的研究及GSE147807数据集）。随后，我们使用相同遗传背景的小鼠品系构建多能干细胞（Pluripotent Stem Cell, PSC），并将其诱导分化为心肌细胞（Cardiomyocyte, CM）。后续通过常规荧光激活细胞分选（FACS）分离得到目标心肌细胞。实验整体设计如下：我们从小鼠体内分离心肌细胞，采样时间点如下（p8及之后的时间点均采用LP-FACS进行分选）：e14、e18、p0、p1、p4、p8、p11、p14、p15、p18、p22、p28、p35、p56及p84。同时我们在以下分化天数收集多能干细胞诱导分化心肌细胞：D8、D10、D12、D15、D18、D25、D30及D45。