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首页> 外文期刊>Cellular and Molecular Bioengineering >Hypertrophy Changes 3D Shape of hiPSC-Cardiomyocytes: Implications for Cellular Maturation in Regenerative Medicine
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Hypertrophy Changes 3D Shape of hiPSC-Cardiomyocytes: Implications for Cellular Maturation in Regenerative Medicine

机译：Hipsc-cardiomyocytes的肥大改变3D形状：对再生医学中的细胞成熟的影响

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Advances in the use of human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes for heart regeneration and in vitro disease models demand a greater understanding of how these cells grow and mature in 3-dimensional space. In this study, we developed an analysis methodology of single cardiomyocytes plated on 2D surfaces to assess their 3D myofilament volume and its z-height distribution, or shape, upon hypertrophic stimulation via phenylephrine (PE) treatment or long-term culture ("aging"). Cardiomyocytes were fixed and labeled with alpha-actinin for confocal microscopy imaging to obtain z-stacks for 3D myofilament volume analysis. In primary neonatal rat ventricular myocytes (NRVMs), area increased 72% with PE, while volume increased 31%. In hiPSC-cardiomyocytes, area increased 70% with PE and 4-fold with aging; however, volume increased significantly only with aging by 2.3-fold. Analysis of z-height myofilament volume distribution in hiPSC-cardiomyocytes revealed a shift from a fairly uniform distribution in control cells to a basally located volume in a more flat and spread morphology with PE and even more so with aging, a shape that was akin to all NRVMs analyzed. These results suggest that 2D area is not a sufficient measure of hiPSC-cardiomyocyte growth and maturation, and that changes in 3D volume and its distribution are essential for understanding hiPSC-cardiomyocyte biology for disease modeling and regenerative medicine applications.
机译：人类诱导多能干细胞（hiPSC）衍生的心肌细胞用于心脏再生和体外疾病模型的研究进展，需要对这些细胞如何在三维空间中生长和成熟有更深入的了解。在这项研究中，我们开发了一种分析方法，对镀在2D表面的单个心肌细胞进行分析，以评估其3D肌丝体积及其z高度分布或形状，通过苯肾上腺素（PE）治疗或长期培养（“老化”）进行肥大刺激。将心肌细胞固定并用α-肌动蛋白标记，用于共焦显微镜成像，以获得用于三维肌丝体积分析的z-叠层。在原代新生大鼠心室肌细胞（NRVM）中，PE使其面积增加72%，而体积增加31%。在hiPSC心肌细胞中，PE使其面积增加70%，随着年龄增长，面积增加4倍；然而，体积仅在老化2.3倍时显著增加。对hiPSC心肌细胞中z高度肌丝体积分布的分析显示，对照细胞中的肌丝体积分布从相当均匀的转变为基底部分布的体积，在PE中表现为更平坦和扩散的形态，在老化中表现得更为明显，这种形态类似于所分析的所有NRVM。这些结果表明，2D面积并不能充分衡量hiPSC心肌细胞的生长和成熟，3D体积及其分布的变化对于理解hiPSC心肌细胞生物学在疾病建模和再生医学中的应用至关重要。

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