Dave Maneesh, Menghini Paola, Sugi Keiki, Somoza Rodrigo A, Lee Zhenghong, Jain Mukesh, Caplan Arnold, Cominelli Fabio
Division of Gastroenterology and Liver Disease, University Hospitals, Digestive Health Research Institute, Case Western Reserve University;
Division of Gastroenterology and Liver Disease, University Hospitals, Digestive Health Research Institute, Case Western Reserve University.
J Vis Exp. 2017 Sep 1(127):55367. doi: 10.3791/55367.
Crohn's disease (CD) is a common chronic inflammatory disease of the small and large intestines. Murine and human mesenchymal stem cells (MSCs) have immunosuppressive potential and have been shown to suppress inflammation in mouse models of intestinal inflammation, even though the route of administration can limit their homing and effectiveness . Local application of MSCs to colonic injury models has shown greater efficacy at ameliorating inflammation in the colon. However, there is paucity of data on techniques to enhance the localization of human bone marrow-derived MSCs (hMSCs) to the small intestine, the site of inflammation in the SAMP-1/YitFc (SAMP) model of experimental Crohn's disease. This work describes a novel technique for the ultrasound-guided intracardiac injection of hMSCs in SAMP mice, a well-characterized spontaneous model of chronic intestinal inflammation. Sex- and age-matched, inflammation-free AKR/J (AKR) mice were used as controls. To analyze the biodistribution and the localization, hMSCs were transduced with a lentivirus containing a triple reporter. The triple reporter consisted of firefly luciferase (fl), for bioluminescent imaging; monomeric red fluorescent protein (mrfp), for cell sorting; and truncated herpes simplex virus thymidine kinase (ttk), for positron emission tomography (PET) imaging. The results of this study show that 24 h after the intracardiac administration, hMSCs localize in the small intestine of SAMP mice as opposed to inflammation-free AKR mice. This novel, ultrasound-guided injection of hMSCs in the left ventricle of SAMP mice ensures a high success rate of cell delivery, allowing for the rapid recovery of mice with minimal morbidity and mortality. This technique could be a useful method for the enhanced localization of MSCs in other models of small-intestinal inflammation, such as TNFΔRE. Future studies will determine if the increased localization of hMSCs by intra-arterial delivery can lead to increased therapeutic efficacy.
克罗恩病（CD）是一种常见的小肠和大肠慢性炎症性疾病。小鼠和人间充质干细胞（MSCs）具有免疫抑制潜能，并且已证实在肠道炎症的小鼠模型中可抑制炎症，尽管给药途径会限制它们的归巢和有效性。将MSCs局部应用于结肠损伤模型已显示出在减轻结肠炎症方面具有更高的疗效。然而，关于增强人骨髓来源的MSCs（hMSCs）在实验性克罗恩病的SAMP-1/YitFc（SAMP）模型中的炎症部位——小肠的定位技术的数据却很匮乏。这项研究描述了一种在SAMP小鼠中超声引导下心内注射hMSCs的新技术，SAMP小鼠是一种特征明确的慢性肠道炎症自发模型。将性别和年龄匹配、无炎症的AKR/J（AKR）小鼠用作对照。为了分析生物分布和定位，用含有三重报告基因的慢病毒转导hMSCs。三重报告基因由萤火虫荧光素酶（fl）组成，用于生物发光成像；单体红色荧光蛋白（mrfp），用于细胞分选；以及截短的单纯疱疹病毒胸苷激酶（ttk），用于正电子发射断层扫描（PET）成像。这项研究的结果表明，心内给药后24小时，hMSCs定位于SAMP小鼠的小肠，而非无炎症的AKR小鼠。这种在SAMP小鼠左心室内进行的新型超声引导hMSCs注射确保了细胞递送的高成功率，使小鼠能够快速恢复，发病率和死亡率降至最低。这项技术可能是一种在其他小肠炎症模型（如TNFΔRE）中增强MSCs定位的有用方法。未来的研究将确定通过动脉内递送增加hMSCs的定位是否能提高治疗效果。
