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首页> 外文期刊>Cell and Tissue Research >Prevascularization of self-organizing engineered heart tissue by human umbilical vein endothelial cells abrogates contractile performance
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Prevascularization of self-organizing engineered heart tissue by human umbilical vein endothelial cells abrogates contractile performance

机译：人脐静脉内皮细胞对自组织的心脏组织的血管形成作用消除了收缩性能

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摘要

Establishing vascularization is a critical obstacle to the generation of engineered heart tissue (EHT) of substantial thickness. Addition of endothelial cells to the formative stages of EHT has been demonstrated to result in prevascularization, or the formation of capillary-like structures. The detailed study of the effects of prevascularization on EHT contractile function is lacking. Here, we evaluated the functional impact of prevascularization by human umbilical vein endothelial cells (HUVECs) in self-organizing EHT. EHT fibers were generated by the self-organization of neonatal rat cardiac cells on a fibrin hydrogel scaffold with or without HUVECs. Contractile function was measured and force-length relationship and rate of force production were assessed. Immunofluorescent studies were used to evaluate arrangement and distribution of HUVECs within the EHT fibers. RT-PCR was used to assess the transcript levels of hypoxia inducible factor-1a (Hif-1a). EHT with HUVECs manifested tubule-like structures at the periphery during fiber formation. After fiber formation, HUVECs were heterogeneously located throughout the EHT fiber and human CD31+ tubule-like structures were identified. The expression level of Hif-1a did not change with the addition of HUVECs. However, maximal force and rate of force generation were not improved in HUVECs containing EHT as compared to control EHT fibers. The addition of HUVECs may result in sparse microvascularization of EHT. However, this perceived benefit is overshadowed by a significant decrease in contractile function and highlights the need for perfused vascularization strategies in order to generate EHT that approaches clinically relevant dimensions.
机译：建立血管化是产生相当厚的工程心脏组织（EHT）的关键障碍。已证明将内皮细胞添加到EHT的形成阶段会导致血管形成前或形成毛细血管样结构。缺乏关于血管形成前对EHT收缩功能影响的详细研究。在这里，我们评估了人脐静脉内皮细胞（HUVEC）在自组织EHT中的血管形成前的功能影响。 EHT纤维是通过在有或没有HUVEC的纤维蛋白水凝胶支架上自组织新生大鼠心脏细胞而产生的。测量收缩功能，评估力长关系和力产生率。免疫荧光研究用于评估EHT纤维中HUVEC的排列和分布。 RT-PCR用于评估缺氧诱导因子-1a（Hif-1a）的转录水平。具有HUVEC的EHT在纤维形成过程中在外围表现出小管状结构。纤维形成后，HUVECs遍布整个EHT纤维，并鉴定出人类CD31 +小管样结构。 Hif-1a的表达水平并没有增加HUVEC。但是，与对照EHT纤维相比，含EHT的HUVEC的最大作用力和产生力的速率没有改善。 HUVEC的添加可能导致EHT的微血管稀疏。但是，这种明显的好处被收缩功能的显着降低所掩盖，并突显了需要灌注血管化策略以产生接近临床相关尺寸的EHT。

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