Name: orthotopic rat kidney transplantation: a novel and simplified surgical approach
Uploaded: 2019-05-07T14:00:00
Description: Watch this Scientific Journal Video about Orthotopic Rat Kidney Transplantation: A Novel and Simplified Surgical Approach at JoVE.com

这项工作的资金来自邦贝克家族庄园的一份慷慨的礼物。

Lewis (RT1 1) 和深色 Agouti (da)(rt1a a) 大鼠是从商业供应商处购买的 (见《材料表》)。这些完全 mhc 不匹配菌株经常被用来研究急性肾移植排斥反应。所有动物都是根据国家卫生研究院 (NIH) 的指导方针, 在约翰·霍普金斯大学的一个特定的无病原体设施中安置和饲养的。所有程序都得到了机构动物护理和使用委员会的批准。
1. 捐助者程序
<ol><li>准备和...

<ol>
	<li>Billingham, R. E., Brent, L., Medawar, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Actively+acquired+tolerance+of+foreign+cells.">Actively acquired tolerance of foreign cells.</a> Nature. 172 (4379), 603-606 (1953).</li><li>Murray, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organ+transplantation+(skin,+kidney,+heart)+and+the+plastic+surgeon.">Organ transplantation (skin, kidney, heart) and the plastic surgeon.</a> Plastic and Reconstructive Surgery. 47 (5), 425-431 (1971).</li><li>Liu, F., Kang, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterotopic+heart+transplantation+in+mice.">Heterotopic heart transplantation in mice.</a> Journal of Visualized Experiments. (6), e238 (2007).</li><li>Ruzza, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterotopic+heart+transplantation+in+rats:+improved+anesthetic+and+surgical+technique.">Heterotopic heart transplantation in rats: improved anesthetic and surgical technique.</a> Transplant Proceedings. 42 (9), 3828-3832 (2010).</li><li>Oldani, G., Lacotte, S., Morel, P., Mentha, G., Toso, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+rats.">Orthotopic liver transplantation in rats.</a> Journal of Visualized Experiments. (65), e4143 (2012).</li><li>Lang, K. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunoprivileged+status+of+the+liver+is+controlled+by+Toll-like+receptor+3+signaling.">Immunoprivileged status of the liver is controlled by Toll-like receptor 3 signaling.</a> Journal of Clinical Investigation. 116 (9), 2456-2463 (2006).</li><li>Sun, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recruitment+of+host+progenitor+cells+in+rat+liver+transplants.">Recruitment of host progenitor cells in rat liver transplants.</a> Hepatology. 49 (2), 587-597 (2009).</li><li>Orlando, G., Soker, S., Wood, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Operational+tolerance+after+liver+transplantation.">Operational tolerance after liver transplantation.</a> Journal of Hepatolgy. 50 (6), 1247-1257 (2009).</li><li>Lodhi, S. 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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nephrotoxicity+of+immunosuppressive+drugs:+long-term+consequences+and+challenges+for+the+future.">Nephrotoxicity of immunosuppressive drugs: long-term consequences and challenges for the future.</a> American Journal of Kidney Diseases. 35 (2), 333-346 (2000).</li><li>Hu, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chimeric+Allografts+Induced+by+Short-Term+Treatment+With+Stem+Cell-Mobilizing+Agents+Result+in+Long-Term+Kidney+Transplant+Survival+Without+Immunosuppression:+A+Study+in+Rats.">Chimeric Allografts Induced by Short-Term Treatment With Stem Cell-Mobilizing Agents Result in Long-Term Kidney Transplant Survival Without Immunosuppression: A Study in Rats.</a> American Journal of Transplantation. 16 (7), 2055-2065 (2016).</li><li>Shrestha, B., Haylor, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+rat+models+of+chronic+allograft+nephropathy:+a+review.">Experimental rat models of chronic allograft nephropathy: a review.</a> International Journal of Nephrology and Renovascular Disease. 7, 315-322 (2014).</li><li>Fu, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Successful+transplantation+of+kidney+allografts+in+sensitized+rats+after+syngeneic+hematopoietic+stem+cell+transplantation+and+fludarabine.">Successful transplantation of kidney allografts in sensitized rats after syngeneic hematopoietic stem cell transplantation and fludarabine.</a> American Journal of Transplantation. 14 (10), 2375-2383 (2014).</li><li>Grau, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immune+Complex-Type+Deposits+in+the+Fischer-344+to+Lewis+Rat+Model+of+Renal+Transplantation+and+a+Subset+of+Human+Transplant+Glomerulopathy.">Immune Complex-Type Deposits in the Fischer-344 to Lewis Rat Model of Renal Transplantation and a Subset of Human Transplant Glomerulopathy.</a> Transplantation. 100 (5), 1004-1014 (2016).</li><li>Vogelbacher, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bortezomib+and+sirolimus+inhibit+the+chronic+active+antibody-mediated+rejection+in+experimental+renal+transplantation+in+the+rat.">Bortezomib and sirolimus inhibit the chronic active antibody-mediated rejection in experimental renal transplantation in the rat.</a> Nephrology Dialysis Transplantation. 25 (11), 3764-3773 (2010).</li><li>Fisher, B., Lee, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microvascular+surgical+techniques+in+research,+with+special+reference+to+renal+transplantation+in+the+rat.">Microvascular surgical techniques in research, with special reference to renal transplantation in the rat.</a> Surgery. 58 (5), 904-914 (1965).</li><li>Mahabir, R. N., Guttmann, R. D., Lindquist, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Renal+transplantation+in+the+inbred+rat.+X.+A+model+of+"weak+histoincompatibility"+by+major+locus+matching.">Renal transplantation in the inbred rat. X. A model of "weak histoincompatibility" by major locus matching.</a> Transplantation. 8 (4), 369-378 (1969).</li><li>Wang, J. J., Hockenheimer, S., Bickerstaff, A. A., Hadley, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+renal+transplantation+procedure.">Murine renal transplantation procedure.</a> Journal of Visualized Experiments. (29), e1150 (2009).</li><li>Bickerstaff, A. A., Wang, J. J., Pelletier, R. P., Orosz, C. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+renal+allografts:+spontaneous+acceptance+is+associated+with+regulated+T+cell-mediated+immunity.">Murine renal allografts: spontaneous acceptance is associated with regulated T cell-mediated immunity.</a> Journal of Immunology. 167 (9), 4821-4827 (2001).</li><li>Silber, S. J., Crudop, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kidney+transplantation+in+inbred+rats.">Kidney transplantation in inbred rats.</a> American Journal of Surgery. 125 (5), 551-553 (1973).</li><li>D'Silva, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+kidney+transplantation+update+with+special+reference+to+vesical+calculi.">Rat kidney transplantation update with special reference to vesical calculi.</a> Microsurgery. 11 (2), 169-176 (1990).</li><li>Yin, M., Booster, M. H., Bogaard, A. E. J. M., Kootstra, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+technique+to+harvest+two+kidneys+from+one+donor+rat+for+transplantation.">A simple technique to harvest two kidneys from one donor rat for transplantation.</a> Lab Animal. 28 (4), 387-390 (1994).</li><li>Lopez-Neblina, F., Toledo-Pereyra, L. H., Suzuki, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrarapid+orthotopic+technique+for+renal+transplantation+in+the+rat.">Ultrarapid orthotopic technique for renal transplantation in the rat.</a> Microsurgery. 15 (4), 274-278 (1994).</li><li>Blom, D., Orloff, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+more+versatile+and+reliable+method+for+renal+transplantation+in+the+rat.">A more versatile and reliable method for renal transplantation in the rat.</a> Microsurgery. 18 (4), 267-269 (1998).</li><li>Engelbrecht, G., Kahn, D., Duminy, F., Hickman, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+rapid+technique+for+renal+transplantation+in+the+rat.">New rapid technique for renal transplantation in the rat.</a> Microsurgery. 13 (6), 340-344 (1992).</li><li>Kline, R., Churchill, M., Churchill, P., Bidani, A., Schwartz, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+osmolality-low+pH+flush+solutions+improve+renal+transplant+function+in+rats.">High osmolality-low pH flush solutions improve renal transplant function in rats.</a> Urology Research. 19 (2), 81-86 (1991).</li><li>Fray, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanism+by+which+renin+secretion+from+perfused+rat+kidneys+is+stimulated+by+isoprenaline+and+inhibited+by+high+perfusion+pressure.">Mechanism by which renin secretion from perfused rat kidneys is stimulated by isoprenaline and inhibited by high perfusion pressure.</a> The Journal of Physiology. 308, 1-13 (1980).</li><li>Fry, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+vascular+endothelial+changes+associated+with+increased+blood+velocity+gradients.">Acute vascular endothelial changes associated with increased blood velocity gradients.</a> Circulation Research. 22 (2), 165-197 (1968).</li><li>Pahlavan, P. S., Smallegange, C., Adams, M. A., Schumacher, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kidney+transplantation+procedures+in+rats:+assessments,+complications,+and+management.">Kidney transplantation procedures in rats: assessments, complications, and management.</a> Microsurgery. 26 (5), 404-411 (2006).</li><li>Savas, C. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Renal+transplantation+in+the+rat--a+new+simple,+non-suture+technique.">Renal transplantation in the rat--a new simple, non-suture technique.</a> Urology Research. 13 (2), 91-93 (1985).</li><li>Fabre, J., Lim, S. H., Morris, P. 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在本手稿中, 我们详细介绍了大鼠原位 KT 的手术方法, 包括执行此过程所需的所有必要设备 (图 5)。1965年, 费舍尔和李发表了第一份关于老鼠 KT 的报告, 这成为一个令人兴奋的调查领域18的开始。自那时以来, 已经引入了许多修改, 以提高该模型的重现性。由于部分和全部 MHC 错配组合的几个自亲和外交系的存在, 为研究缺血再灌注损伤和肾移植排斥反应和耐?...

从历史上看, 第一次移植排斥研究是由布伦特和医疗通过在啮齿类动物1中进行皮肤移植进行的。很快就可以看出, 皮肤具有明显的免疫学特征, 使其成为一个高度免疫原性器官, 不同于其他血管化固体器官2的排斥反应。大鼠对固体器官移植排斥反应的研究通常仅限于心脏、肝脏和肾脏移植。虽然这些器官中的每一个都适合研究排斥, 但每个器官都有优点和缺点。心脏移植通常移植到腹部, 并与主动脉和静脉吻合, 受者的本地心脏就位 3.这并不能重现人类的临床、解剖和生理状况。此外, 心脏对冷缺血非常敏感, 必须在1小时内优先再灌注, 才能恢复其功能4。肝脏移植通常被认为是手术更具挑战性和时间敏感性的执行。去除原生肝脏后, 捐献者的肝脏必须在30分钟内植入和再灌注, 因为没有正常运作的肝脏,受者就无法持续更长的时间5。肝动脉、门静脉, 尤其是胆管重建需要精细的手术技巧。除了手术的挑战, 肝脏是已知的具有耐受性的属性和啮齿类动物和人类可以成为操作耐受6,7,8。肾脏, 不像前面提到的器官, 可以移植在一个原位的方式, 已知是一个免疫原器官与一致的, 可重复的排斥发作 (如果不是免疫抑制), 并允许长时间的冷缺血时间小时。这使得大鼠肾移植成为研究同种异体移植排斥反应和耐受性的理想模型。
肾移植 (KT) 是终末期肾病患者的首选治疗方法。在过去几十年中, KT 之后的短期生存结果有了显著改善, 但长期生存结果却停滞不前。常规免疫抑制方案仍然是标准的抗排斥治疗。然而, 长期使用免疫抑制疗法会导致显著的发病率和死亡率, 如肾毒性、糖尿病和10、11、12只继发性恶性肿瘤.长期, 慢性抗体和脂肪介导的排斥威胁移植物的生存, 有限的治疗选择可用。
移植的一个主要目标是诱导移植耐受性, 以避免慢性免疫抑制的需要。大鼠 KT 模型是研究免疫排斥过程和评估免疫调节和移植耐受新方法的可靠工具。大鼠也是研究急性和慢性细胞和抗体介导的排斥13,14,15,16, 17的合适模型。这种手术模型已被证明是一个可靠的, 可重复的, 和具有成本效益的工具, 以研究同种异体移植排斥和耐受的各个方面。在进行昂贵而繁琐的大型动物研究之前, 它经常被用来测试新的公差诱导方案。在大鼠身上进行 KT 手术需要广泛的手术培训和专业知识, 才能达到 gt;90% 的存活率。在这份手稿和随附的教学视频中, 我们为大鼠的原位 KT 提供了一个逐步的大纲, 在我们的机构成功地执行了多年。
在开始任何程序之前, 捐献者和受赠者的选择都是至关重要的, 这取决于实验的性质。理想情况下, 捐献者和受赠者的体重应在220-260 克之间, 年龄在8-12周之间。220克以下的动物有小直径的动脉、静脉和输尿管, 这使得接受者的吻合特别具有挑战性。轻微的失血会导致低血容量, 并导致较小的动物死亡。体重超过260克的动物在血管周围显示出更多的脂肪, 血管隔离将需要更多的手术时间, 并增加冷缺血时间。

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	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Buprenorphine HCL</td>
			<td style="width:167px;">Reckitt Benckiser Healthcare UK</td>
			<td style="width:167px;">NDC12496-0757-5</td>
			<td style="width:463px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Dissecting forceps, curved</td>
			<td style="width:167px;">Zhenbang, China</td>
			<td style="width:167px;"></td>
			<td style="width:463px;">11cm Flat handle&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Heparin sodium injection USP</td>
			<td style="width:167px;">Sagent Pharmaceuticals&#160;</td>
			<td style="width:167px;">NDC25021-400-10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Micro-forceps, straight, smooth</td>
			<td style="width:167px;">Jingzhong, China</td>
			<td style="width:167px;">WA3010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Micro needle holder</td>
			<td style="width:167px;">Jingzhong, China</td>
			<td style="width:167px;">WA2010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Micro vessel clamps</td>
			<td style="width:167px;">Jingzhong, China</td>
			<td style="width:167px;">WA40120</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Micro spring sciccor 1</td>
			<td style="width:167px;">ROBOZ</td>
			<td style="width:167px;">RS-5620</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Micro spring sciccor 2</td>
			<td style="width:167px;">F.S.T.</td>
			<td style="width:167px;">91501-09</td>
			<td></td>
		</tr>
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			<td height="21" style="height:21px;width:216px;">Micro spring sciccor 3</td>
			<td style="width:167px;">Zhenbang, China</td>
			<td style="width:167px;"></td>
			<td style="width:463px;">8.5cm Vannas,curved</td>
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			<td height="21" style="height:21px;width:216px;">Prograf (Tacrolimus/FK506)</td>
			<td style="width:167px;">Astellas</td>
			<td style="width:167px;"></td>
			<td></td>
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			<td height="42" style="height:42px;width:216px;">Rats</td>
			<td style="width:167px;">Charles River &#38; Taconic Biosciences&#160;</td>
			<td style="width:167px;">LEW/Crl &#38; DA-M&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Shaver</td>
			<td style="width:167px;">Wahl</td>
			<td>79600-2101</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Suture 4-0</td>
			<td style="width:167px;">Ethicon</td>
			<td style="width:167px;">J304H</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Suture, 4-0&#160;</td>
			<td style="width:167px;">Ethicon</td>
			<td style="width:167px;">683G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Suture, 10-0&#160;</td>
			<td style="width:167px;">Ethicon</td>
			<td style="width:167px;">2820G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Syringes &#38; Needles</td>
			<td style="width:167px;">BD</td>
			<td style="width:167px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Thread, 8-0</td>
			<td style="width:167px;">Ashaway</td>
			<td style="width:167px;">75290</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ureteral cuff</td>
			<td style="width:167px;">Microlumen</td>
			<td style="width:167px;">160-1</td>
			<td>Polymide Tubing, Diameter 0.41 mm&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Venous cuff</td>
			<td style="width:167px;">Intramedic BD</td>
			<td style="width:167px;">7441</td>
			<td>PE-200 Non-radiopaque polyethylene tubing ID: 1.4 mm, OD: 1.9 mm</td>
		</tr>
	</tbody>
</table>

orthotopic rat kidney transplantation: a novel and simplified surgical approach

与任何类型的肾脏替代疗法相比, 肾移植为终末期肾病患者提供了更高的生存率和更高的生活质量。在过去的几十年里, 大鼠肾移植模型被用来研究排斥和耐受的免疫现象。在进行昂贵的临床前大型动物研究之前, 该模型已成为测试新的免疫调节药物和方案的不可或缺的工具。
该方案提供了如何可靠地在大鼠中进行原位肾移植的详细概述。该方案包括三个独特的步骤, 增加成功的概率: 通过冲洗门静脉灌注供肾和使用袖系统来吻合肾静脉和输尿管, 从而减少寒冷和温暖缺血时间。使用这种技术, 我们已经达到了存活率超过6个月与正常血清肌酐的动物与同种或耐受的肾脏移植。根据研究的目的, 这种模型可以通过移植前或移植后的治疗来改变, 以研究急性、慢性、细胞或抗体介导的排斥反应。研究肾移植的不同方面是一种可重复、可靠、经济高效的动物模型。

我们进行了同源 (n = 5) 和同种异体肾移植 (n = 5)。进行同源移植的动物在没有任何免疫抑制治疗的情况下实现了长期生存。在没有免疫抑制的情况下接受同种异体移植的动物拒绝了移植, 并死于肾功能衰竭, 平均生存期为 8天 (图 4a)。在同种组中, 平均血清肌酐略有增加, 而同种组则增加了 14倍 (0.5 mg/dL 对 7.0 mg/dl, p & lt;0.01) (图 4<...

Watch this Scientific Journal Video about Orthotopic Rat Kidney Transplantation: A Novel and Simplified Surgical Approach at JoVE.com

Orthotopic Rat Kidney Transplantation: A Novel and Simplified Surgical Approach

本手稿和程序的目的是详细解释和演示大鼠原位肾移植的手术方法。该方法简化后, 采用静脉和输尿管袖吻合术, 实现供肾的正确灌注, 缩短再灌注时间。

大鼠原位肾移植: 一种新的简化的手术方法

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of ...

我们在约翰霍普金斯大学的实验室一直在进行小动物肾脏和肝移植超过20年的经验。此特定协议概述了在大鼠中成功进行正畸肾移植所需的关键步骤。为了增加成功的概率和可重复性，我们的协议包括三个独特的步骤，通过门户静脉灌注供体肾脏，并使用袖口系统对肾脏静脉和输尿管进行抗剖。演示这个程序的将是乐琪博士，一个研究生在我们的实验室。要开始此过程，请对捐赠大鼠进行麻醉，如文本协议所述。将大鼠放在一个苏皮位置，用无菌遮蔽胶带固定四肢。应用眼部润滑剂，对手术场进行消毒，如文本协议中所述。在第一次切口之前，通过检查没有头趾捏戒反射，确保大鼠被充分麻醉。使用剪刀，首先做一个大的纵向中线皮肤和肌肉切口从交响乐到西腓，并进入腹腔。用潮湿的无菌纱布盖住肠道，并转移到腹部的右侧，露出大肛门、维纳卡瓦和左肾。用一毫升的cc注射器涂抹一毫升预热盐水，以保持肠道和腹部器官的湿润和正常温度。应用第二个湿润的纱布覆盖和调动胃和脾颅到肾脏。然后添加一个小的湿纱布，以覆盖暴露的肾脏。使用显微手术解剖钳，从结缔组织和相互分离和调动左肾动脉和静脉。现在通过烧灼左肾静脉来分离左肾静脉，通过烧灼性肾内动脉来隔离左肾动脉。之后，通过解剖的钳子解剖结缔组织，调动大动脉和维纳卡瓦的优越和低于左肾的骨骼。使用解剖钳子从结缔组织中分割并调动尿管，然后使用微型剪刀在从肾脏骨盆中测量大约两厘米的长度进行对角切口。将聚酰胺袖口插入尿尿器的一半，通过放置 8-0 的结来固定袖口丝绸缝合线。使用解剖钳或微型剪刀将其从肾上腺脂肪分离来调动左肾。保留肾脏附着的脂肪胶囊，并使用该站点处理肾脏。现在，使用注射器通过笔静脉用27针施用200个单位肝素。用棉签对注射部位施加压力至少一分钟，以防止出血。识别门户静脉和劣质的维纳卡瓦。暴露劣等的维娜卡瓦。使用 16 表针将 50 毫升与 500 单位肝素混合在门户静脉中，准备冲洗肾脏。冲洗前，在红外线水平上切割劣质静脉注射，在针头插入位点切开入口静脉，让血液排出血液循环。开始冲洗肾脏逐渐注入盐水溶液。观察肾脏的颜色从深红色到均匀的灰色或淡色。冲洗后，将肾动脉和静脉近到主动脉和静脉注射。将冲洗过的肾脏放在含有冷盐水的培养皿中，然后将捐赠者的身体从手术场中移走。一旦肾脏在冷盐水中，固定和固定静脉袖口的手柄，并轻轻地拉下袖口的肾静脉。然后使用 8 - 0 放置三节， 将肾静脉放在袖口上丝绸缝合线。要开始接收程序，请重复从捐赠程序到肝素管理的步骤。然后将两个创伤微瓣夹放在左肾动脉和静脉近到主动脉和静脉卡瓦。将接受者肾静脉近到肾脏的入口。用肝素盐水冲洗肾静脉，将所有剩余的血液从血管中排出。将连接的肾静脉滑过以前位于供体肾脏的袖口肾静脉。然后用 8 - 0 固定静脉丝绸缝合线。在将肾静脉固定到袖口上时，保持相同的位置方向。将肾动脉近到接受者肾脏的入口。用肝素盐水冲洗，去除血管中多余的血液。现在将尿尿器在左肾的下极水平。从肾上脂肪中调动肾脏，取出原生肾脏。使用 10-0 尼龙缝合线，用 8-10 个中断的缝合线对肾动脉进行端到端的动脉瘤。使用冒险层操纵动脉。拆下容器夹以重新启动肾脏的再灌注。首先拆下静脉上的夹子，然后取下动脉上的夹子。使用无菌棉签为动脉瘤病区周围的任何渗出区域增加轻压。几分钟应该足以获得专利的动脉瘤。简要观察肾脏，以评估足够的灌注。重新灌注后，肾脏应立即改变颜色，几分钟后逐渐恢复其自然深红色。有时观察到尿尿和现场尿液生产的可见近位。将尿套的外露尖端插入接收器尿尿器，然后用 8-0 固定接收器尿修器。丝绸领带。为了保持供体和接收器的位置，将每个尿尿器的末端彼此系在一起。可选地，用3-0丝线来绑下肾骨，使右肾切除。从腹腔中去除所有纱布，将所有器官返回其自然位置。然后喷一毫升盐水在肠道上，以保持他们潮湿。在直肠肌肉上使用 4-0 可吸收的缝合线和 4-0 丝缝合线关闭腹部，以中断的方式关闭皮肤层。将动物放在一个干净的笼子里，可以接触到饮用水和食物，并允许在37摄氏度的加热垫上恢复。观察恢复一到两个小时，然后再将动物放回动物设施。患有联合肾移植的动物无需任何免疫抑制治疗即可获得长期生存。相反，接受异体肾移植而不进行免疫抑制的动物则拒绝移植，以8天的中位存活期死于肾衰竭。平均血清肌氨酸在合成组中适度增加，而在异基因组增加14倍。切除后，合成肾的宏观视图均未出现任何异常。肾的颜色和内部结构仍然完好无损。相比之下，被拒动物的肾脏所有图谱呈现红色出血斑块与内部结构的破坏。血氧林和Eeosin的合成移植物的染色显示薄球状毛细血管循环与正常数量的内皮细胞和中分细胞。拒绝所有显示被破坏的球状结构与炎症和管状炎的迹象。CD8阳性染色，以确认T细胞中培养排斥。虽然合成全图显示很少CD8阳性T细胞，但被拒绝的所有图谱显示，在球状细胞和图巴利周围，CD8阳性细胞的数量显著增加，证实了T细胞的抑制。该模型为排斥、耐受和供体-宿主相互作用的免疫学现象提供了重要见解，为固体器官移植治疗性临床干预的发展铺平道路。

Research

JoVE Journal

Medicine

Orthotopic Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy

主动脉原位移植大鼠模型，研究慢性血管病变的发展

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant ...

科研

医学

A Novel Surgical Approach for Intratracheal Administration of Bioactive Agents in a Fetal Mouse Model

一种新的手术方法气管内注射生物活性剂在胎鼠模型

Prenatal pulmonary delivery of cells, genes or pharmacologic agents could provide the basis for new therapeutic strategies for a variety of genetic and ...

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

部分原位肝移植大鼠模型的肝动脉重建的外科手术

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

小鼠体外循环模型: 一种新方法

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization ...

A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction

应用新型磁吻合技术 Suprahepatic 腔静脉重建大鼠原位肝移植模型的研究

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep ...

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

正交心脏移植的临床前猪模型

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart ...

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

矫形肝移植大鼠模型中动脉重联后并发症减少

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...

Orthotopic Kidney Auto-Transplantation in a Porcine Model Using 24 Hours Organ Preservation And Continuous Telemetry

使用24小时器官保存和连续遥测的猪模型中正交肾自动移植

In the present era of organ transplantation with critical organ shortage, various strategies are employed to expand the pool of available allografts for ...

Robot-Assisted Kidney Transplantation

机器人辅助肾移植

This paper describes robot-assisted kidney transplantation (RAKT) from a living donor. The robot is docked between the parted legs of the patient, placed ...