Roche Christopher D, Gentile Carmine
The University of Sydney; University of Technology Sydney (UTS); The Royal North Shore Hospital; University Hospital of Wales.
The University of Sydney; University of Technology Sydney (UTS); The Royal North Shore Hospital;
J Vis Exp. 2020 Sep 26(163). doi: 10.3791/61675.
Testing regenerative properties of 3D bioprinted cardiac patches in vivo using murine models of heart failure via permanent left anterior descending (LAD) ligation is a challenging procedure and has a high mortality rate due to its nature. We developed a method to consistently transplant bioprinted patches of cells and hydrogels onto the epicardium of an infarcted mouse heart to test their regenerative properties in a robust and feasible way. First, a deeply anesthetized mouse is carefully intubated and ventilated. Following left lateral thoracotomy (surgical opening of the chest), the exposed LAD is permanently ligated and the bioprinted patch transplanted onto the epicardium. The mouse quickly recovers from the procedure after chest closure. The advantages of this robust and quick approach include a predicted 28-day mortality rate of up to 30% (lower than the 44% reported by other studies using a similar model of permanent LAD ligation in mice). Moreover, the approach described in this protocol is versatile and could be adapted to test bioprinted patches using different cell types or hydrogels where high numbers of animals are needed to optimally power studies. Overall, we present this as an advantageous approach which may change preclinical testing in future studies for the field of cardiac regeneration and tissue engineering.
通过永久性左前降支（LAD）结扎使用心力衰竭小鼠模型在体内测试3D生物打印心脏补片的再生特性是一项具有挑战性的程序，因其性质导致死亡率很高。我们开发了一种方法，以稳健且可行的方式将生物打印的细胞和水凝胶补片一致地移植到梗死小鼠心脏的心外膜上，以测试其再生特性。首先，将深度麻醉的小鼠小心插管并通气。在左外侧开胸（胸部手术开口）后，将暴露的LAD永久性结扎，并将生物打印的补片移植到心外膜上。小鼠在胸部闭合后很快从手术中恢复。这种稳健且快速的方法的优点包括预测的28天死亡率高达30%（低于其他使用类似小鼠永久性LAD结扎模型的研究报告的44%）。此外，本方案中描述的方法具有通用性，可适用于使用不同细胞类型或水凝胶测试生物打印补片，在这种情况下需要大量动物来优化研究效能。总体而言，我们将此作为一种有利的方法，它可能会改变未来心脏再生和组织工程领域的临床前测试。
