Nora Eccles Harrison Cardiovascular Research and Training Institute, University of Utah, Salt Lake City, Utah, USA.
InviRX Antiviral Therapeutics, Sandy, Utah, USA.
Cell Physiol Biochem. 2024 Oct 4;58(5):538-547. doi: 10.33594/000000730.
BACKGROUND/AIMS: Advances in induced pluripotent stem cell (iPSC) technology allow for reprogramming of adult somatic cells into stem cells from which patient- and disease-specific cardiomyocytes (CMs) can be derived. Yet, the potential of iPSC technology to revolutionize cardiovascular research is limited, in part, by the embryonic nature of these cells. Here, we test the hypothesis that decellularized porcine left ventricular extracellular cardiac matrix (ECM) provides environmental cues that promote transcriptional maturation and patterning of iPSC-CMs in culture.
Cardiac progenitor cells were plated on ECM or standard tissue plates (2D monolayer) for 30 days, after which CM orientation and single cell transcriptomics were evaluated using confocal imaging and singe cell RNA-sequencing, respectively.
Cardiac progenitors differentiated on left ventricular ECM formed longitudinal fibers that differed quantitatively from progenitors differentiated in standard 2D conditions. Unsupervised clustering of single cell transcriptomics identified a CM cluster expressing a higher level of genes related to CM maturation. CMs differentiated on ECM were overrepresented in this cluster, indicating a bias toward CM maturation, compared to cells differentiated in standard 2D monolayer conditions.
Our data suggest that environmental cues related to the left ventricular ECM may promote differentiation to a more mature CM state compared to cells differentiated on a standard 2D monolayer, while facilitating organization into longitudinal micro-fibers. Our study highlights the utility of ECM as a differentiation substrate to promote CM maturation and fiber orientation .
背景/目的：诱导多能干细胞（iPSC）技术的进步允许将成人体细胞重编程为多能干细胞，从中可以衍生出患者特异性和疾病特异性心肌细胞（CM）。然而，iPSC 技术在心血管研究中具有潜在的革命性，部分原因是这些细胞的胚胎性质。在这里，我们测试了这样一个假设，即脱细胞化的猪左心室细胞外心脏基质（ECM）提供了环境线索，可促进 iPSC-CM 在培养中的转录成熟和模式形成。
将心脏祖细胞种植在 ECM 或标准组织板（2D 单层）上 30 天，然后使用共聚焦成像分别评估 CM 取向和单细胞转录组学。
在左心室 ECM 上分化的心脏祖细胞形成了纵向纤维，其在数量上与在标准 2D 条件下分化的祖细胞不同。单细胞转录组学的无监督聚类鉴定出一个表达更高水平与 CM 成熟相关基因的 CM 簇。与在标准 2D 单层条件下分化的细胞相比，在 ECM 上分化的 CM 在该簇中过度表达，表明 CM 成熟存在偏向性。
我们的数据表明，与左心室 ECM 相关的环境线索可能比在标准 2D 单层上分化的细胞更能促进向更成熟的 CM 状态分化，同时促进组织成纵向微纤维。我们的研究强调了 ECM 作为一种分化底物的实用性，可促进 CM 成熟和纤维取向 。
