Biochemical and Biophysical Research Communications ( IF 2.5 ) Pub Date : 2020-12-02 , DOI: 10.1016/j.bbrc.2020.11.076 Xiao Ni , Ke Xu , Yunfei Zhao , Jingyan Li , Linli Wang , Fengxu Yu , Guang Li
Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) play a critical role in most translational and clinical applications. Although glucose starvation (GS) has been evaluated during cellular purification, there has been no comprehensive evaluation of the transcriptional heterogeneity of these cells. Here, we applied GS for 3 days starting at day 10 of differentiation, and then, harvested hiPSC-CMs at day 20 for single-cell RNA sequencing (scRNA-seq). We found that GS dramatically reduced the proportion of non-cardiomyocytes cells and increased the number of late-stage cardiomyocytes. We also recorded an increase in the expression of MYH6, MYH7, ACTN2, TNNT2, and several other genes associated with the structural and functional maturation of cardiomyocytes. Further analysis indicated that these changes were focused on the signaling pathways involved in the regulation of the actin cytoskeleton, cardiac muscle development, and cardiac muscle contraction. Finally, pseudotime analysis revealed that GS hiPSC-CMs developed in a more mature direction. Together, these results suggest that GS treatment improves the purity and maturation of hiPSC-CMs, which should increase the feasibility of hiPSC-CMs applications.
中文翻译:
单细胞分析揭示了hiPSC-CM中葡萄糖饥饿的纯化和成熟作用
人类诱导的多能干细胞来源的心肌细胞(hiPSC-CM)在大多数翻译和临床应用中都起着至关重要的作用。尽管已经在细胞纯化过程中评估了葡萄糖饥饿(GS),但尚未全面评估这些细胞的转录异质性。在这里,我们从分化的第10天开始应用GS 3天,然后在第20天收获hiPSC-CM用于单细胞RNA测序(scRNA-seq)。我们发现GS大大减少了非心肌细胞的比例,并增加了晚期心肌细胞的数量。我们还记录了MYH6,MYH7,ACTN2,TNNT2和与心肌细胞的结构和功能成熟相关的其他几个基因的表达增加。进一步的分析表明,这些变化集中在肌动蛋白细胞骨架,心肌发育和心肌收缩的调控中。最后,伪时间分析表明,GS hiPSC-CMs向更成熟的方向发展。总之,这些结果表明GS处理提高了hiPSC-CM的纯度和成熟度,这应该增加hiPSC-CM应用的可行性。