颈椎异位心脏移植小鼠
Summary
The cuff technique shortens and facilitates the model of heterotopic cervical heart transplantation in mice by avoiding technically challenging suture anastomoses of small vessels. The technical details shown in this video paper should allow researches to establish this model in their laboratories.
Abstract
The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies this model by avoiding challenging suture anastomoses of small vessels thereby reducing warm ischemia time. In comparison to abdominal graft implantation the cervical model is less invasive and the implanted graft is easily accessible for further follow-up examinations. Anastomoses are performed by pulling the ascending aorta of the graft over the cuff with the recipient’s common carotid artery and by pulling the main pulmonary artery over the cuff with the external jugular vein. Selection of appropriate cuff size and complete mobilization of the vessels are important for successful revascularization. Ischemia-reperfusion (I/R) injury can be minimized by perfusing the graft with a cardioplegic solution and by hypothermia. In this article, we provide technical details for a simplified and improved cuff technique, which should allow surgeons with basic microsurgical skills to perform the procedure with a high success rate.
Introduction
在心脏移植模型经常用于研究I / R损伤，移植排斥反应，移植后血管病变和免疫调节剂的效率。小鼠模型结合许多优点，如公知的免疫学和遗传背景，许多近交和转基因品系和相对低的成本的实验的可用性。
1973年，腹部心脏移植小鼠的技术由科里等 [1]首次描述。在此模型中的移植物是由升主动脉到主肺动脉到下腔静脉的收信人的腹主动脉和的端 – 侧吻合术吻合血管。 1991年，宫颈心脏移植小鼠缝合技术由陈等人 2被描述。在此模型中一个逆行冠状动脉灌注由收件人的颈动脉的吻合建立到克的升主动脉船尾。经由冠状窦静脉血液排入右心房，右心室，并通过肺动脉到受体颈外静脉（图1）排出。相较于腹腔移植植入颈椎模型是创伤小，移植物的位置表浅能够很容易获得额外的随访检查（ 如触诊，超声心动图，活体荧光显微镜）3,4。
尽管技术改进5,6，更广泛地使用这种模式已被有限的，由于技术上的困难进行了小血管缝合吻合术吻合口瘘和血栓形成的高发期。 1991年，松浦晃一郎等人 。引入其中血管壁被外翻在一个合成柱面，大大方便了颈部心脏移植7的模型的袖口技术。该技术是福rther通过了各种实验移植模型，包括8肾，胰9，四肢10，11,12肝，肺13,14和血管移植15。相比于缝合技术吻合术的所需时间，因此，热缺血时间较短，有显著较少血管并发症使用袖套法16。此外，它使医生与显微稍加培训与成功率高执行该操作。袖带技术的缺点是在颈总动脉，这可能会改变脑灌注的前提结扎。
在这个视频文章中，我们介绍了宫颈心脏移植小鼠改进和简化套管法。
Protocol
Representative Results
Discussion
我们已经完成了超过200个心脏移植，成功率为90％，在各种实验设置。与这里所描述的技术中，完整的程序，可在60分钟内由有经验的外科医生进行。由器官获取之前制备颈部血管，总接枝缺血时间可以限制到20分钟。时间用于植入（热缺血时间）是显著缩短使用袖套法相比，缝合技术16。
而不是简单地切除如前所述5,17的心脏，我们通过灌注移植与临床使用的?…
Divulgaciones
The authors have nothing to disclose.
Acknowledgements
M.-A. Deutsch is supported by Dr. Rusche Forschungsprojekt (2014) Deutsche Stiftung für Herzforschung and Deutsche Gesellschaft für Thorax- Herz- und Gefäßchirurgie. M. Krane is supported by Deutsche Stiftung für Herzforschung (F/37/11), Deutsches Zentrum für Herz Kreislauf Forschung (DZHK B 13-050A; DZHK B 14-013SE) and Deutsche Forschungsgemeinschaft (KR3770/7-1; KR3770/9-1)
Materials
| Forceps JFC-7.2 | S&T | P-00036 | Curved tip 0.20 mm |
| Vessel dilatator D-5a | S&T | S-00124 | |
| Adventita scissor | S&T | S-00102 | |
| Vascular clamp B-1V | S&T | S-00396 | |
| Yasargil Clip | Peter Lazic | 65,097 | 3.5 mm lenght |
| Bipolar forceps | Micromed | 148-100-011 | tip 0.25 mm |
| Polyimide tubes | River Tech Medical | 19-24 gauge | |
| 8.0 silk ligatures | Catgut | ||
| Custodiol HTK solution | Essential Pharmaceuticals |
Referencias
- Corry, R. J., Winn, H. J., Russell, P. S. Heart transplantation in congenic strains of mice. Transplant Proc. 5, 733-735 (1973).
- Chen, Z. H. A technique of cervical heterotopic heart transplantation in mice. Transplantation. 52, 1099-1101 (1991).
- Martins, P. N. Assessment of graft function in rodent models of heart transplantation. Microsurgery. 28, 565-570 (2008).
- Schramm, R., et al. The subepicardial microcirculation in heterotopically transplanted mouse hearts: an intravital multifluorescence microscopy study. J Thorac Cardiovasc Surg. 134, 210-217 (2007).
- Liu, F., Kang, S. M. Heterotopic heart transplantation in mice. J Vis Exp : JoVE. 238, (2007).
- Gong, W., et al. Introduction of modified cervical cardiac transplant model in mice. Exp Clin Transplant. 10, 158-162 (2012).
- Matsuura, A., Abe, T., Yasuura, K. Simplified mouse cervical heart transplantation using a cuff technique. Transplantation. 51, 896-898 (1991).
- Chen, H., Zhang, Y., Zheng, D., Praseedom, R. K., Dong, J. Orthotopic kidney transplantation in mice: technique using cuff for renal vein anastomosis. PloS one. 8, e77278 (2013).
- Maglione, M., et al. A novel technique for heterotopic vascularized pancreas transplantation in mice to assess ischemia reperfusion injury and graft pancreatitis. Surgery. 141, 682-689 (2007).
- Sucher, R., et al. Orthotopic hind-limb transplantation in rats. J Vis Exp : JoVE. , (2010).
- Kamada, N., Calne, R. Y. Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage. Transplantation. 28, 47-50 (1979).
- Oldani, G., Lacotte, S., Morel, P., Mentha, G., Toso, C. Orthotopic liver transplantation in rats. J Vis Exp : JoVE. , (2012).
- Okazaki, M., et al. A mouse model of orthotopic vascularized aerated lung transplantation. Am J Transplant. 7, 1672-1679 (2007).
- Suzuki, H., Fan, L., Wilkes, D. S. Development of obliterative bronchiolitis in a murine model of orthotopic lung transplantation. J Vis Exp : JoVE. , (2012).
- Zou, Y., et al. Mouse model of venous bypass graft arteriosclerosis. Am J Pathol. 153, 1301-1310 (1998).
- Zhou, Y., Gu, X., Xiang, J., Qian, S., Chen, Z. A comparative study on suture versus cuff anastomosis in mouse cervical cardiac transplant. Exp Clin Transplant. 8, 245-249 (2010).
- Niimi, M. The technique for heterotopic cardiac transplantation in mice: experience of 3000 operations by one surgeon. J Heart Lung Transplant. 20, 1123-1128 (2001).
- Arras, M., Autenried, P., Rettich, A., Spaeni, D., Rulicke, T. Optimization of intraperitoneal injection anesthesia in mice: drugs, dosages, adverse effects, and anesthesia depth. Comp Med. 51, 443-456 (2001).
