Conventional rigid 2D substrates cause complex contractile signals in monolayers of human induced pluripotent stem cell-derived cardiomyocytes.
Conventional rigid 2D substrates cause complex contractile signals in monolayers of human induced pluripotent stem cell-derived cardiomyocytes.
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DOI：
10.1113/jp282228
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发表时间：
2022-03
影响因子：
5.5
通讯作者：
Smith, Godfrey L.
中科院分区：
文献类型：
--
作者：
Huethorst, Eline;Mortensen, Peter;Simitev, Radostin D.;Gao, Hao;Pohjolainen, Lotta;Talman, Virpi;Ruskoaho, Heikki;Burton, Francis L.;Gadegaard, Nikolaj;Smith, Godfrey L.
关键词：
单层中的人诱导多能干细胞衍生的心肌细胞（hiPSC-CM）通过细胞-细胞和细胞-基质粘附进行机械相互作用。在hiPSC-CM单层中分析收缩的时空特征：（1）附着于玻璃或塑料（杨氏模量（E）> 1 GPa），（2）分离（无基质）和（3）附着于柔性胶原水凝胶（E = 22 kPa）。比较了异丙肾上腺素对刚性和柔性基质收缩的影响。为了阐明潜在的机制，进行了进一步的基因表达和计算研究。HiPSC-CM单层在刚性表面上表现出多相收缩特征，而水凝胶、无基质培养物或单细胞仅观察到简单的抽搐样时程。异丙肾上腺素没有改变任何一个表面上的收缩曲线，但其lusotropic和变时作用在水凝胶中比在玻璃中更大。在应激激活基因NPPA和NPPB的表达方面，刚性和柔性基底之间没有显著差异。细胞簇的计算模型表明，由于细胞与细胞的功能异质性，在刚性基底上存在类似的复杂收缩相互作用。刚性生物材料表面引起hiPSC-CM单层的非生理性多相收缩。柔性基质对于正常的抽搐样收缩动力学和变力性干预的解释是必要的。 对接种在常规刚性表面（玻璃或塑料）上的人诱导多能干细胞衍生心肌细胞（hiPSC-CM）单层的时空收缩性分析显示，尽管在相同区域记录的动作电位相同，但整个单层存在具有高度变异性的多相收缩模式。这些多相模式不存在于单细胞、分离的单层细胞或接种在软基质（如水凝胶）上的单层细胞中，其中仅观察到"抽搐"样瞬变。显示高百分比区域具有多相收缩的HiPSC-CM单层具有显著增加的收缩持续时间和降低的亲膜性药物反应。没有迹象表明多相收缩模式与应激激活的NPPA或NPPB信号通路的显著激活相关。细胞簇的计算模型支持以下生物学发现：刚性表面和差异细胞-基质粘附是hiPSC-CM多相收缩行为的基础。 对接种在常规刚性表面（玻璃或塑料）上的人诱导多能干细胞衍生的心肌细胞（hiPSC-CM）单层的时空收缩性分析显示，尽管动作电位相同，但单层上的可变多相收缩事件。这些多相模式不存在于单细胞、分离的单层细胞或接种在软基质（如水凝胶）上的单层细胞中，其中仅观察到生理性"抽搐"样瞬变。细胞簇的计算模型支持的生物学发现，部分粘附细胞簇的刚性表面导致多相收缩行为。实验数据表明，与柔性基底上的反应相比，刚性基底上的hiPSC-CM单层具有显著增加的收缩持续时间和降低的亲葡萄糖药物反应。尽管刚性表面上的收缩事件具有非生理性质，但没有迹象表明多相模式与应激激活信号通路的显著较高激活相关。
Human induced pluripotent stem cell‐derived cardiomyocytes (hiPSC‐CM) in monolayers interact mechanically via cell–cell and cell–substrate adhesion. Spatiotemporal features of contraction were analysed in hiPSC‐CM monolayers (1) attached to glass or plastic (Young's modulus (E) >1 GPa), (2) detached (substrate‐free) and (3) attached to a flexible collagen hydrogel (E = 22 kPa). The effects of isoprenaline on contraction were compared between rigid and flexible substrates. To clarify the underlying mechanisms, further gene expression and computational studies were performed. HiPSC‐CM monolayers exhibited multiphasic contractile profiles on rigid surfaces in contrast to hydrogels, substrate‐free cultures or single cells where only simple twitch‐like time‐courses were observed. Isoprenaline did not change the contraction profile on either surface, but its lusitropic and chronotropic effects were greater in hydrogel compared with glass. There was no significant difference between stiff and flexible substrates in regard to expression of the stress‐activated genes NPPA and NPPB. A computational model of cell clusters demonstrated similar complex contractile interactions on stiff substrates as a consequence of cell‐to‐cell functional heterogeneity. Rigid biomaterial surfaces give rise to unphysiological, multiphasic contractions in hiPSC‐CM monolayers. Flexible substrates are necessary for normal twitch‐like contractility kinetics and interpretation of inotropic interventions. Spatiotemporal contractility analysis of human induced pluripotent stem cell‐derived cardiomyocyte (hiPSC‐CM) monolayers seeded on conventional, rigid surfaces (glass or plastic) revealed the presence of multiphasic contraction patterns across the monolayer with a high variability, despite action potentials recorded in the same areas being identical. These multiphasic patterns are not present in single cells, in detached monolayers or in monolayers seeded on soft substrates such as a hydrogel, where only ‘twitch’‐like transients are observed. HiPSC‐CM monolayers that display a high percentage of regions with multiphasic contraction have significantly increased contractile duration and a decreased lusotropic drug response. There is no indication that the multiphasic contraction patterns are associated with significant activation of the stress‐activated NPPA or NPPB signalling pathways. A computational model of cell clusters supports the biological findings that the rigid surface and the differential cell–substrate adhesion underly multiphasic contractile behaviour of hiPSC‐CMs. Abstract figure legend Spatio‐temporal contractility analysis of human induced pluripotent stem cell‐derived cardiomyocyte (hiPSC‐CM) monolayers seeded on conventional rigid surfaces (glass or plastic) showed variable multiphasic contraction events across the monolayer despite identical action potentials. These multiphasic patterns are not present in single cells, in detached monolayers or in monolayers seeded on soft substrates such as a hydrogel, where only physiological ‘twitch’‐like transients are observed. A computational model of cell clusters supports the biological findings that partial adhesion of cell clusters to a rigid surface causes multiphasic contractile behaviour. Experimental data showed that hiPSC‐CM monolayers on rigid substrates have significantly increased contractile duration and a decreased lusotropic drug response when compared to responses on a flexible substrate. Despite the unphysiological nature of the contractile events on rigid surfaces, there is no indication that the multiphasic patterns are associated with significantly higher activation of the stress‐activated signalling pathways.