Contractility and Calcium Transient Maturation in the Human iPSC-Derived Cardiac Microfibers.
Contractility and Calcium Transient Maturation in the Human iPSC-Derived Cardiac Microfibers.
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DOI：
10.1021/acsami.2c07326
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发表时间：
2022-08-10
影响因子：
9.5
通讯作者：
Yang, Huaxiao
中科院分区：
文献类型：
--
作者：
Strimaityte, Dovile;Tu, Chengyi;Yanez, Apuleyo;Itzhaki, Ilanit;Wu, Haodi;Wu, Joseph C.;Yang, Huaxiao
关键词：
人诱导的多能干细胞衍生的心肌细胞（hiPSC-CM）被认为在肌节组织、收缩机制、钙瞬变和转录组谱方面不成熟，这阻止了它们在建模和研究心脏发育和疾病中的进一步应用。为了提高 hiPSC-CM 的成熟度，我们通过基于模板的微图案化方法将 hiPSC-CM 设计成心脏微纤维（iCMF），这使得 hiPSC-CM 能够以端到端连接方式排列，以便在生理硬度的水凝胶上长时间培养。采用一系列表征方法来评估 iCMF 在结构和功能水平上的成熟程度，包括免疫组织化学、钙瞬态、逆转录定量 PCR、心肌收缩力和电起搏分析。我们的结果表明，与微图案或随机单个 hiPSC-CM 以及相同细胞数 iCMF 中随机簇中的 hiPSC-CM 相比，iCMF 中 hiPSC-CM 的心脏成熟度有所改善。我们发现肌节长度增加，肌节有更好的规律性和排列，收缩性增强，钙瞬变成熟，T 管形成，粘附连接和间隙连接形成改善。 iCMF 中的 hiPSC-CM 显示出强大的钙循环，以响应 0.5 至 7 Hz 的编程和连续电起搏。此外，我们利用肌球蛋白结合蛋白 C (MYBPC3) 突变的 hiPSC-CM 生成了 iCMF，以验证 iCMF 在模拟心脏肥厚表型方面的概念。这些发现表明，hiPSC-CM 的多模式 iCMF 连接在结构和功能上促进了心脏的成熟，这将揭示 hiPSC-CM 模型在心肌病和心脏再生医学疾病建模中应用的全部潜力。
Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are considered immature in the sarcomere organization, contractile machinery, calcium transient, and transcriptome profile, which prevent them from further applications in modeling and studying cardiac development and disease. To improve the maturity of hiPSC-CMs, here we engineered the hiPSC-CMs into cardiac microfibers (iCMFs) by a stencil-based micropatterning method, which enables the hiPSC-CMs to be aligned in an end-to-end connection for prolonged culture on the hydrogel of physiological stiffness. A series of characterization approaches were performed to evaluate the maturation in iCMFs on both structural and functional levels, including immunohistochemistry, calcium transient, reverse-transcription quantitative PCR, cardiac contractility, and electrical pacing analysis. Our results demonstrate an improved cardiac maturation of hiPSC-CMs in iCMFs compared to the micropatterned or random single hiPSC-CMs, and hiPSC-CMs in a random cluster at the same cell number of iCMFs. We found an increased sarcomere length, better regularity and alignment of sarcomeres, enhanced contractility, matured calcium transient, and T-tubule formation, and improved adherens junction and gap junction formation. The hiPSC-CMs in iCMFs showed a robust calcium cycling in response to the programmed and continuous electrical pacing from 0.5 to 7 Hz. Moreover, we generated the iCMFs with hiPSC-CMs with mutations in myosin-binding protein C (MYBPC3) to have a proof-of-concept of iCMFs in modeling cardiac hypertrophic phenotype. These findings suggest that the multipatterned iCMF connection of hiPSC-CMs boosts the cardiac maturation structurally and functionally, which will reveal the full potential of the application of hiPSC-CM models in disease modeling of cardiomyopathy and cardiac regenerative medicine.
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影响因子：
14
作者：
Rao, Christopher;Prodromakis, Themistoklis;Kolker, Ljudmila;Chaudhry, Umar A. R.;Trantidou, Tatiana;Sridhar, Arun;Weekes, Claire;Camelliti, Patrizia;Harding, Sian E.;Darzi, Ara;Yacoub, Magdi H.;Athanasiou, Thanos;Terracciano, Cesare M.
通讯作者：
Terracciano, Cesare M.
DOI：
10.1073/pnas.1508073112
发表时间：
2015-10-13
影响因子：
11.1
作者：
Ribeiro, Alexandre J. S.;Ang, Yen-Sin;Pruitt, Beth L.
通讯作者：
Pruitt, Beth L.
DOI：
10.1016/j.bbamcr.2015.11.005
发表时间：
2016-07
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Bedada FB;Wheelwright M;Metzger JM
通讯作者：
Metzger JM
影响因子：
8
作者：
Helms, Adam S.;Tang, Vi T.;Day, Sharlene M.
通讯作者：
Day, Sharlene M.
影响因子：
20.1
作者：
Angst, BD;Khan, LUR;Gourdie, RG
通讯作者：
Gourdie, RG