Department of Cardiovascular Surgery, University Heart and Vascular Center Hamburg, Hamburg, Germany.
DZHK (German Centre for Cardiovascular Research), partner site Hamburg/Kiel/Lübeck, Hamburg, Germany.
Artif Organs. 2024 Nov;48(11):1251-1263. doi: 10.1111/aor.14833. Epub 2024 Jul 23.
Engineered Heart Tissue (EHT) is a promising tool to repair heart muscle defects and can additionally be used for drug testing. Due to the absence of an in vitro vascularization, EHT geometry crucially impacts nutrient and oxygen supply by diffusion capacity. We analyzed cardiomyocyte survival in different EHT geometries.
Different geometries with varying surface-area-to-volume-ratios were calculated (structure A (Ring) AS/V = 58.47 mm/440 μL, structure B (Infinity) 25.86 mm/440 μL). EHTs were generated from hiPSC-derived cardiomyocytes (4 × 10) and a fibrin/thrombin hydrogel. Cell viability was evaluated by RT-PCR, cytometric studies, and Bioluminescence imaging.
Using 3D-printed casting molds, spontaneously beating EHTs can be generated in various geometric forms. At day 7, the RT-PCR analyses showed a significantly higher Troponin-T value in ring EHTs, compared to infinity EHTs. In cytometric studies, we evaluated 15% more Troponin-T positive cells in ring (73% ± 12%), compared to infinity EHTs (58% ± 11%, p = 0.04). BLI visualized significantly higher cell survival in ring EHTs (ROI = A: 1.14 × 10 p/s and B: 8.47 × 10 p/s, p < 0.001) compared to infinity EHTs during longitudinal cultivation process.
Use of 3D-printing allows the creation of EHTs in all desired geometric shapes. The geometry with an optimized surface-area-to-volume-ratio (ring EHT) demonstrated a significantly higher cell survival measured by RT-PCR, Bioluminescence imaging, and cytometric studies using FACS analysis.
工程心脏组织（EHT）是修复心肌缺陷的有前途的工具，并且还可以用于药物测试。由于缺乏体外血管化，EHT 几何形状通过扩散能力对营养物质和氧气供应产生至关重要的影响。我们分析了不同 EHT 几何形状中的心肌细胞存活率。
计算了具有不同表面积与体积比的不同几何形状（结构 A（环）AS/V=58.47mm/440μL，结构 B（无限）25.86mm/440μL）。EHT 由 hiPSC 衍生的心肌细胞（4×10）和纤维蛋白/凝血酶水凝胶生成。通过 RT-PCR、细胞计量学研究和生物发光成像评估细胞活力。
使用 3D 打印铸造模具，可以在各种几何形状中生成自发跳动的 EHT。在第 7 天，RT-PCR 分析显示，与无限 EHT 相比，环 EHT 的肌钙蛋白-T 值显著更高。在细胞计量学研究中，我们评估了环 EHT 中阳性细胞的肌钙蛋白-T 增加了 15%（73%±12%），而无限 EHT 为 58%±11%（p=0.04）。BLI 可视化显示，在纵向培养过程中，环 EHT 中的细胞存活率显著更高（ROI=A：1.14×10p/s 和 B：8.47×10p/s，p<0.001）。
3D 打印的使用允许在所有所需的几何形状中创建 EHT。具有优化表面积与体积比的几何形状（环 EHT）在通过 RT-PCR、生物发光成像和使用 FACs 分析的细胞计量学研究中显示出更高的细胞存活率。
