Givens Sophie E, Andebrhan Abygail A, Wang Ruchen, Kong Xiangzhen, Rothermel Taylor M, Hosseini Sanaz, Xie An, Shameem Mohammad, Torniainen Andrea A, Ebrahimi-Barough Somayeh, Boland Samuel F, Johnson Maya, Mancipe Natalia Calixto, Singh Bhairab N, Dudley Samuel, Alford Patrick W, Tolkacheva Elena G, van Berlo Jop H, Ogle Brenda M
Biomedical Engineering, University of Minnesota, Minneapolis, MN, USA.
Lillehei Heart Institute (LHI), Department of Medicine, University of Minnesota, Minneapolis, MN, USA.
Stem Cell Reports. 2025 Aug 12;20(8):102572. doi: 10.1016/j.stemcr.2025.102572. Epub 2025 Jul 3.
Accumulating evidence indicates that maturation limits cardiomyocyte proliferation. We expand on that theory by co-culturing human induced pluripotent stem cell (hiPSC)-cardiomyocytes (CM) with epicardial cells (EPCs) and epicardial-derived cells in both 2D co-cultures and 3D engineered heart tissues (EHTs). In 2D co-cultures, the percentage of proliferating CM increased in parallel with stark electrophysiologic improvements. Single-cell transcriptomics revealed a significant shift in the bulk CM population of the epicardial-CM co-cultures as characterized by more fetal-like myofilament isoforms but with enhanced pathways associated with electrochemical maturation. The 3D-EHTs containing EPCs showed more limited proliferation but a similar improvement in CM electrophysiologic function. Next, epicardial-derived fibroblasts (EPD-FBs) were added to the EHTs containing EPCs, and we observed significant myofilament maturation and increased force generation. Our results suggest that some aspects of CM maturation (i.e., electrochemical) can occur when proliferation rates are relatively high, and that sarcomere-associated mechanical maturation occurs at later developmental stages when proliferation has largely ceased.
越来越多的证据表明，成熟会限制心肌细胞增殖。我们通过在二维共培养和三维工程心脏组织（EHT）中将人诱导多能干细胞（hiPSC）-心肌细胞（CM）与心外膜细胞（EPC）和心外膜衍生细胞共培养，对该理论进行了拓展。在二维共培养中，增殖性CM的百分比随着明显的电生理改善而平行增加。单细胞转录组学显示，心外膜-CM共培养的大量CM群体发生了显著变化，其特征是更多胎儿样肌丝异构体，但与电化学成熟相关的途径增强。含有EPC的三维EHT显示出更有限的增殖，但CM电生理功能有类似改善。接下来，将心外膜衍生的成纤维细胞（EPD-FB）添加到含有EPC的EHT中，我们观察到明显的肌丝成熟和力产生增加。我们的结果表明，CM成熟的某些方面（即电化学方面）可以在增殖率相对较高时发生，而肌节相关的机械成熟发生在发育后期，此时增殖已基本停止。
