Human Induced Pluripotent Stem Cell Encapsulation Geometry Impacts Three-Dimensional Developing Human Engineered Cardiac Tissue Functionality
Human Induced Pluripotent Stem Cell Encapsulation Geometry Impacts Three-Dimensional Developing Human Engineered Cardiac Tissue Functionality
复制标题
人类诱导多能干细胞封装几何形状影响三维发育的人类工程心脏组织功能
DOI：
10.1089/ten.tea.2022.0107
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发表时间：
2022
影响因子：
4.1
通讯作者：
Young, Nathan P.
中科院分区：
文献类型：
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作者：
Ellis, Morgan E.;Harris, Bryana N.;Hashemi, Mohammadjafar;Harvell, B. Justin;Bush, Michaela Z.;Hicks, Emma E.;Finklea, Ferdous B.;Wang, Eric M.;Nataraj, Ravikiran;Young, Nathan P.
几十年来，心脏组织工程一直致力于减轻心血管疾病的巨大负担。为了改善心脏组织的均匀性和心肌细胞（CM）的成熟，在本研究中，我们研究了在三维（3D）直接心脏分化平台上改变初始包封几何形状。传统的工程化心脏组织生产是利用预分化的CMs来产生三维心脏组织，通常涉及各种细胞选择和外源刺激方法来促进CM成熟。直接用人类诱导多能干细胞（hiPSCs）而不是预分化的CMs开始组织形成，简化了工程心脏组织形成过程，使其更适用于广泛实施和规模化。在这项研究中，hiPSCs被聚乙二醇纤维蛋白原包裹在三种组织几何形状（圆盘状微岛、正方形和矩形）中，并进行既定的心脏分化方案。随着时间的推移，每种几何形状的3D工程心脏组织（3D- ects）显示出相似的CM群体（约65%）和基因表达。值得注意的是，矩形组织显示出较少的组织异质性，并显示出成熟CMs的更多高级特征，包括肌纤维排列和z线形成。矩形组织的各向异性收缩性能显著高于正方形和微岛状组织（MI 0.28±0.03,SQ 0.35±0.05,RT 0.79±0.04）。这项研究展示了一种简化和改进3D-ECT生产的直接方法，无需使用外源性机械或电起搏，并且有可能用于生物打印和药物测试应用。目前改善心脏成熟后分化的方法仍然繁琐和复杂。在这项研究中，我们研究了初始封装几何形状对改善三维工程心脏组织（3D-ECT）生产和三种组织几何形状（包括盘状微岛、正方形和矩形）分化后成熟的影响。值得注意的是，与微岛组织和正方形组织相比，矩形3D-ECTs显示出较少的组织异质性和更多成熟心肌细胞的高级特征，包括肌纤维排列、z线形成和各向异性收缩特性。这项研究表明，最初的人类诱导多能干细胞包裹的矩形组织几何结构可以促进心脏成熟，而不是实施细胞选择或繁琐的分化后操作，包括外源性机械和/或电起搏。
Cardiac tissue engineering has been working to alleviate the immense burden of cardiovascular disease for several decades. To improve cardiac tissue homogeneity and cardiomyocyte (CM) maturation, in this study, we investigated altering initial encapsulation geometry in a three-dimensional (3D) direct cardiac differentiation platform. Traditional engineered cardiac tissue production utilizes predifferentiated CMs to produce 3D cardiac tissue and often involves various cell selection and exogenous stimulation methods to promote CM maturation. Starting tissue formation directly with human induced pluripotent stem cells (hiPSCs), rather than predifferentiated CMs, simplifies the engineered cardiac tissue formation process, making it more applicable for widespread implementation and scale-up. In this study, hiPSCs were encapsulated in poly (ethylene glycol)-fibrinogen in three tissue geometries (disc-shaped microislands, squares, and rectangles) and subjected to established cardiac differentiation protocols. Resulting 3D engineered cardiac tissues (3D-ECTs) from each geometry displayed similar CM populations (∼65%) and gene expression over time. Notably, rectangular tissues displayed less tissue heterogeneity and suggested more advanced features of maturing CMs, including myofibrillar alignment and Z-line formation. In addition, rectangular tissue showed significantly higher anisotropic contractile properties compared to square and microisland tissues (MI 0.28 ± 0.03, SQ 0.35 ± 0.05, RT 0.79 ± 0.04). This study demonstrates a straightforward method for simplifying and improving 3D-ECT production without the use of exogenous mechanical or electrical pacing and has the potential to be utilized in bioprinting and drug testing applications.Impact statementCurrent methods for improving cardiac maturation postdifferentiation remain tedious and complex. In this study, we examined the impact of initial encapsulation geometry on improvement of three-dimensional engineered cardiac tissue (3D-ECT) production and postdifferentiation maturation for three tissue geometries, including disc-shaped microislands, squares, and rectangles. Notably, rectangular 3D-ECTs displayed less tissue heterogeneity and more advanced features of maturing cardiomyocytes, including myofibrillar alignment, Z-line formation, and anisotropic contractile properties, compared to microisland and square tissues. This study demonstrates an initial human induced pluripotent stem cell-encapsulated rectangular tissue geometry can improve cardiac maturation, rather than implementing cell selection or tedious postdifferentiation manipulation, including exogenous mechanical and/or electrical pacing.