Conradi Lenard, Pahrmann Christiane, Schmidt Stephanie, Deuse Tobias, Hansen Arne, Eder Alexandra, Reichenspurner Hermann, Robbins Robert C, Eschenhagen Thomas, Schrepfer Sonja
Transplant and Stem Cell Immunobiology Lab and CVRC, University Hospital Hamburg, University Heart Center Hamburg.
J Vis Exp. 2011 Jun 1(52):2605. doi: 10.3791/2605.
Various techniques of cardiac tissue engineering have been pursued in the past decades including scaffolding strategies using either native or bioartificial scaffold materials, entrapment of cardiac myocytes in hydrogels such as fibrin or collagen and stacking of myocyte monolayers. These concepts aim at restoration of compromised cardiac function (e.g. after myocardial infarction) or as experimental models (e.g. predictive toxicology and substance screening or disease modelling). Precise monitoring of cell survival after implantation of engineered heart tissue (EHT) has now become possible using in-vivo bioluminescence imaging (BLI) techniques. Here we describe the generation of fibrin-based EHT from a transgenic rat strain with ubiquitous expression of firefly luciferase (ROSA/luciferase-LEW Tg; ). Implantation is performed into the greater omentum of different rat strains to assess immune responses of the recipient organism following EHT implantation. Comparison of results generated by BLI and the Enzyme Linked Immuno Spot Technique (ELISPOT) confirm the usability of BLI for the assessment of immune responses.
在过去几十年里,人们探索了多种心脏组织工程技术,包括使用天然或生物人工支架材料的支架策略、将心肌细胞包埋在诸如纤维蛋白或胶原蛋白的水凝胶中以及堆叠心肌细胞单层。这些概念旨在恢复受损的心脏功能(如心肌梗死后)或作为实验模型(如预测毒理学和物质筛选或疾病建模)。现在,使用体内生物发光成像(BLI)技术可以精确监测工程心脏组织(EHT)植入后的细胞存活情况。在这里,我们描述了从萤火虫荧光素酶普遍表达的转基因大鼠品系(ROSA/荧光素酶-LEW Tg)生成基于纤维蛋白的EHT。将其植入不同大鼠品系的大网膜中,以评估EHT植入后受体生物体的免疫反应。BLI和酶联免疫斑点技术(ELISPOT)产生的结果比较证实了BLI在评估免疫反应方面的可用性。