Chen Xiaojuan, Kaufman Dixon B
Division of Organ Transplantation, Department of Surgery, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA.
Methods Mol Biol. 2009;574:75-85. doi: 10.1007/978-1-60327-321-3_7.
Bioluminescence imaging (BLI) modalities have been developed, refined, and used broadly in the study of small animal models of human biology and disease, including monitoring the fate of transplanted islets in vivo in real time. In order to advance our understanding of the pathophysiology and immunobiology of islet transplantation as they occur in living animals, islet grafts tagged with light-emitting luciferase can be implanted in a mouse islet transplantation model and assessed using in vivo BLI. We have utilized transgenic islets expressing the firefly luciferase as donor islets in syngeneic and allogeneic islet transplant mouse models for monitoring islets in vivo by BLI after they have been transplanted at different sites of the mice, including the intrahepatic site via portal vein injection. The sensitive and non-invasive BLI system allows better understanding of the dynamic fate of transplanted islets and the relationships among the islet mass that ultimately engrafts, the quality of graft function, and overall glucose homeostasis. It permits detection of early changes in islet graft function or mass due to rejection to prompt timely therapeutic intervention and change the fate of the graft. This chapter details some of the procedures for islet isolation, transplantation, and imaging as well as considerations of using the BLI system in the field of islet transplantation research.
生物发光成像(BLI)技术已得到发展、完善,并广泛应用于人类生物学和疾病的小动物模型研究,包括实时监测体内移植胰岛的命运。为了加深我们对活体动物胰岛移植病理生理学和免疫生物学的理解,可将标记有发光荧光素酶的胰岛移植到小鼠胰岛移植模型中,并使用体内BLI进行评估。我们已在同基因和异基因胰岛移植小鼠模型中,利用表达萤火虫荧光素酶的转基因胰岛作为供体胰岛,通过BLI在小鼠不同部位(包括经门静脉注射至肝内部位)移植后监测体内胰岛。灵敏且非侵入性的BLI系统有助于更好地了解移植胰岛的动态命运,以及最终植入的胰岛质量、移植功能质量和整体葡萄糖稳态之间的关系。它能够检测由于排斥反应导致的胰岛移植功能或质量的早期变化,从而促使及时进行治疗干预并改变移植的命运。本章详细介绍了胰岛分离、移植和成像的一些程序,以及在胰岛移植研究领域使用BLI系统的注意事项。