Armentero Marie-Therese, Bossolasco Patrizia, Cova Lidia
Laboratory of Functional Neurochemistry, C. Mondino National Institute of Neurology Foundation, IRCCS, Pavia, Italy.
Methods Mol Biol. 2013;1052:13-28. doi: 10.1007/7651_2013_21.
The recently developed near-infrared (NIR) light imaging technology combines low background noise with deep tissue penetration and readily allows imaging and tracking of NIR-labeled cells, following transplantation in small animal model of diseases. The real-time and longitudinal detection of grafted cells in vivo, as well as their rapid ex vivo localization, may further clarify graft interactions with the surrounding, in target and nontarget organs throughout the body, over time. The present chapter describes a protocol for (1) the efficient labeling of human mesenchymal stem cells (hMSCs) using a membrane intercalating dye, emitting in the NIR 815 nm spectrum; (2) the stereotaxic transplantation of NIR 815-hMSCs in rodent model of Parkinson's disease; and (3) the longitudinal in vivo detection of the grafted cells and the subsequent ex vivo imaging in selected tissues.
最近开发的近红外（NIR）光成像技术结合了低背景噪声和深层组织穿透能力，在疾病的小动物模型中移植后，可以轻松地对近红外标记的细胞进行成像和追踪。体内移植细胞的实时和纵向检测以及它们在体外的快速定位，可能会随着时间的推移进一步阐明移植细胞与全身靶器官和非靶器官周围环境的相互作用。本章描述了一个方案，用于（1）使用在近红外815nm光谱中发射的膜插入染料对人间充质干细胞（hMSCs）进行有效标记；（2）在帕金森病啮齿动物模型中对近红外815-hMSCs进行立体定向移植；以及（3）对移植细胞进行纵向体内检测，并在选定组织中进行后续体外成像。
