小鼠肾脏移植技术
Summary
The goal of this manuscript is to describe the steps required to perform a kidney transplant in a mouse, paying particular attention to the details of the arterial anastomosis.
Abstract
The first mouse kidney transplant technique was published in 19731 by the Russell laboratory. Although it took some years for other labs to become proficient in and utilize this technique, it is now widely used by many laboratories around the world. A significant refinement to the original technique using the donor aorta to form the arterial anastomosis instead of the renal artery was developed and reported in 1993 by Kalina and Mottram 2 with a further advancement coming from the same laboratory in 1999 3. While one can become proficient in this model, a search of the literature reveals that many labs still experience a high proportion of graft loss due to arterial thrombosis. We describe here a technique that was devised in our laboratory that vastly reduces the arterial thrombus reported by others 4,5. This is achieved by forming a heel-and-toe cuff of the donor infra-renal aorta that facilitates a larger anastomosis and straighter blood flow into the kidney.
Introduction
自1973年以来在小鼠肾脏移植模型一直是一个有价值的研究工具，但技术问题阻碍了它的广泛使用。多年来，一些论文已发表的细节改进/改进这一程序。作为主要血管实体器官移植模型这个过程可能是仅次于异位心脏移植模型，这也是在1973年6设计由罗素实验室。这两种型号借给自己的研究分为异体排斥反应，对移植肾功能延迟发展和局部缺血再灌注损伤。
其中最常见的问题与肾移植报是动脉血栓形成4,5,7，我们也经历了我们实验室的发病率相对较高。因此，我们开始着手进行血栓形成的文献综述，并可能发现这个技术问题的原因，也设计一个可能的解决方案。血栓形成的最可能的原因是血液需要从收件人主动脉，成体供肾肾主动脉，然后就到了供肾动脉有点曲折的道路。此路径将导致湍流，肾动脉，可导致血小板激活和血栓形成。根据近期的观察和搜索相关文献8-14中 ，我们提出了一个新的技术，减少血栓形成，以0％。
此处所描述的技术从在动脉踵和胎趾袖口哪个设施改善血液流的形成以前报道的技术而不同，显著减少血栓形成。袖带是通过将横跨肾动脉口的面的红外线肾主动脉在低于45°到主动脉（图1A＆1B）的纵向轴线的角度形成的。这导致了袖带大约2mm的长度。静脉卡雷尔补丁是由横断第r形成烯醛静脉进入下腔静脉，从而增加了封套的直径。该红外供肾腹主动脉脚跟和脚趾袖口端 – 端吻合于受体腹主动脉和供体肾静脉/下腔静脉修补程序端 – 端吻合给收件人腹部下腔静脉（IVC） 。输尿管随后被引入并如由韩等人3锚固在膀胱。
在这项研究中未经处理的移植，只有热缺血时间（ 即 ，不冷缺血）进行比较。在这种情况下，热缺血是指从血液流过供体肾脏的停止的时间（下面的步骤1.11），并在受者（下面步骤2.11）的接枝的再灌注。冷缺血是指肾脏未灌注和保存在冷藏直到植入程序开始的时间。
Protocol
Representative Results
Discussion
掌握这种移植技术是困难的，但一旦完成了它是一个非常强有力的研究工具。病人的外科医生/研究人员将通过注重细节和技术，这是在小动物模型的关键是掌握任何外科手术，更使一致性得到回报。掌握鼠标肾移植的技术难度大很多倍，而且极有可能在其他小动物移植模型中的经验，必须处理这个过程之前获得。
即，将针以保持一个专利管腔时的相对的血管壁的…
Divulgations
The authors have nothing to disclose.
Acknowledgements
这项工作是由1R03DK096151的部分资助。
Materials
| Instrument | Roboz # | Fine Science Tools # | Arosurgical # |
| Straight micro-dissecting forcep #5 | RS-5015 | 11295-51 | |
| Curved micro-dissecting forcep #7 | RS-5047 | 11297-00 | |
| Curved serrated forcep | RS-5137 | 11052-10 | |
| Vannas micro-dissecting scissors, short | RS-5610 | 09.140.08 | |
| Micro-dissecting scissors, straight, sharp, long | 11.602.11 | ||
| Micro spring handle needle holder | 11.549.15 | ||
| Straight mosquito forcep | 91308-12 | ||
| Micro-dissecting scissors, straight, blunt | RS-5962 | 14078-10 | |
| Micro-dissecting scissors, curved, blunt | RS-5981 | 14079-10 | |
| Micro retractor | RS-6540 | ||
| Instrument tray, 10” x 6 ½” x ¾” | RT-1350S | ||
| Silk suture, 5/0, 22.5m spool | 18020-50 | ||
| Suture | |||
| 10/0 nylon | T4A10Q07 | ||
| 5/0 silk | E19A05N | ||
| Gloves | Drapes | ||
| Biogel from Medex Supply | Precept, #64-9012-9 | ||
| Syringes | Cotton applicators | ||
| B-D 1cc insulin, #329424 | Fisher-brand, #23-400-100 | ||
| Povidone-Iodine swabs | |||
| PDI, #B40600 | |||
| 4/0 Cotton ties | |||
| Domestic cotton autoclaved with instruments | |||
References
- Skoskiewicz, M., Chase, C., Winn, H. J., Russell, P. S. Kidney transplants between mice of graded immunogenetic diversity. Transplant Proc. 5 (1), 721-725 (1973).
- Kalina, S. L., Mottram, P. L. A microsurgical technique for renal transplantation in mice. Aust N Z J Surg. 63 (3), 213-216 (1993).
- Han, W. R., Murray-Segal, L. J., Mottram, P. L. Modified technique for kidney transplantation in mice. Microsurgery. 19 (6), 272-274 (1999).
- Ge, F., Gong, W. Strategies for successfully establishing a kidney transplant in a mouse model. Exp Clin Transplant. 9 (5), 287-294 (2011).
- Martins, P. N. Learning curve, surgical results and operative complications for kidney transplantation in mice. Microsurgery. 26 (8), 590-593 (2006).
- Corry, R. J., Winn, H. J., Russell, P. S. Primarily vascularized allografts of hearts in mice. The role of H-2D, H-2K, and non-H-2 antigens in rejection. Transplantation. 16 (4), 343-350 (1973).
- Zhang, Z., Chen, H., Qian, S. Kidney Transplantation in Mice. Experimental Organ Transplantation. , 45-64 (2013).
- Zhang, L., Moskovitz, M., Piscatelli, S., Longaker, M. T., Siebert, J. W. Hemodynamic study of different angled end-to-side anastomoses. Microsurgery. 16 (2), 114-117 (1995).
- Liu, Q., Mirc, D., Fu, B. M. Mechanical mechanisms of thrombosis in intact bent microvessels of rat mesentery. J Biomech. 41 (12), 2726-2734 (2008).
- Chesnutt, J. K., Han, H. C. Tortuosity triggers platelet activation and thrombus formation in microvessels. J Biomech Eng. 133 (12), 121004 (2011).
- Han, H. C. Blood vessel buckling within soft surrounding tissue generates tortuosity. J Biomech. 42 (16), 2797-2801 (2009).
- Gutierrez-Diaz, J. A., et al. Intraluminal thrombus and neointimal hyperplasia after microvascular surgery. Surg Neurol. 24 (2), 153-159 (1985).
- Khouri, R. K., Cooley, B. C., Kenna, D. M., Edstrom, L. E. Thrombosis of microvascular anastomoses in traumatized vessels: fibrin versus platelets. Plast Reconstr Surg. 86 (1), 110-117 (1990).
- Johnson, P. C., et al. Initial platelet deposition at the human microvascular anastomosis: effect on downstream platelet deposition to intact and injured vessels. Plast Reconstr Surg. 90 (4), 650-658 (1992).
