Name: a modified surgical technique for kidney transplantation in mice
Uploaded: 2022-07-22T13:15:00
Description: Watch this Scientific Journal Video about A Modified Surgical Technique for Kidney Transplantation in Mice at JoVE.com

我们感谢白天天医生团队在画外音方面的帮助，感谢包勉小姐在医学插画方面的帮助。这项工作得到了德国研究基金会（DFG）的部分支持，以促进国际合作（HO2581 / 4-1至AH）和中国国家科学基金（NSFC;#81760291至FJ）。

所有动物实验均根据欧洲议会关于保护用于科学目的的动物的指令2010/63 / EU的指南进行（动物伦理卡：下萨克森州食品和药物安全部，#33.9-42502-04-11/0492）。使用无菌手术器械和耗材（高压灭菌）进行手术，并尽量保持手术区域无菌。
注意：C57BL / 6J雄性小鼠作为供体和受体（同源移植模型），而Balb / c小鼠作为肾脏同种异体移植受体（研究急性同种异体移植排斥模型<sup class="x...

<ol>
	<li>Skoskiewicz, M., Chase, C., Winn, H. J., Russell, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kidney+transplants+between+mice+of+graded+immunogenetic+diversity.">Kidney transplants between mice of graded immunogenetic diversity.</a> Transplantation Proceedings. 5 (1), 721-725 (1973).</li><li>Jiang, K., 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=Noninvasive+assessment+of+renal+fibrosis+with+magnetization+transfer+MR+imaging:+Validation+and+evaluation+in+murine+renal+artery+stenosis.">Noninvasive assessment of renal fibrosis with magnetization transfer MR imaging: Validation and evaluation in murine renal artery stenosis.</a> Radiology. 283 (1), 77-86 (2017).</li><li>Tse, G. H., 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=Mouse+kidney+transplantation:+Models+of+allograft+rejection.">Mouse kidney transplantation: Models of allograft rejection.</a> Journal of Visualized Experiments. (92), e52163 (2014).</li><li>Okazaki, 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=et+al.Costimulatory+blockade-mediated+lung+allograft+acceptance+is+abrogated+by+overexpression+of+Bcl-2+in+the+recipient.">et al.Costimulatory blockade-mediated lung allograft acceptance is abrogated by overexpression of Bcl-2 in the recipient.</a> Transplantation Proceedings. 41 (1), 385-387 (2009).</li><li>Chuck, N. C., 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=et+al.Ultra-short+echo-time+magnetic+resonance+imaging+distinguishes+ischemia/reperfusion+injury+from+acute+rejection+in+a+mouse+lung+transplantation+model.">et al.Ultra-short echo-time magnetic resonance imaging distinguishes ischemia/reperfusion injury from acute rejection in a mouse lung transplantation model.</a> Transplant International. 29 (1), 108-118 (2016).</li><li>Zhang, 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=Pattern+of+liver,+kidney,+heart,+and+intestine+allograft+rejection+in+different+mouse+strain+combinations.">Pattern of liver, kidney, heart, and intestine allograft rejection in different mouse strain combinations.</a> Transplantation. 62 (9), 1267-1272 (1996).</li><li>Wang, 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>Plenter, R., Jain, S., Ruller, C. M., Nydam, T. L., Jani, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+kidney+transplant+technique.">Murine kidney transplant technique.</a> Journal of Visualized Experiments. (104), e52848 (2015).</li><li>Plenter, R. J., Jain, S., Nydam, T. L., Jani, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revised+arterial+anastomosis+for+improving+murine+kidney+transplant+outcomes.">Revised arterial anastomosis for improving murine kidney transplant outcomes.</a> Journal of Investigative Surgery. 28 (4), 208-214 (2015).</li><li>Rong, S., Lewis, A. G., Kunter, U., Haller, H., Gueler, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+knotless+technique+for+kidney+transplantation+in+the+mouse.">A knotless technique for kidney transplantation in the mouse.</a> Journal of Transplantation. , 127215 (2012).</li><li>Kreimann, K., 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=Ischemia+reperfusion+injury+triggers+CXCL13+release+and+B-cell+recruitment+after+allogenic+kidney+transplantation.">Ischemia reperfusion injury triggers CXCL13 release and B-cell recruitment after allogenic kidney transplantation.</a> Frontiers in Immunology. 11, 1204 (2020).</li><li>Schmidbauer, 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=Diffusion-Weighted+imaging+and+mapping+of+T1+and+T2+relaxation+time+for+evaluation+of+chronic+renal+allograft+rejection+in+a+translational+mouse+model.">Diffusion-Weighted imaging and mapping of T1 and T2 relaxation time for evaluation of chronic renal allograft rejection in a translational mouse model.</a> Journal of Clinical Medicine. 10 (19), 4318 (2021).</li><li>Wu, K., 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=Novel+technique+for+blood+circuit+reconstruction+in+mouse+heart+transplantation+model.">Novel technique for blood circuit reconstruction in mouse heart transplantation model.</a> Microsurgery. 26, 594-598 (2006).</li><li>Haas, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+allograft+nephropathy+or+interstitial+fibrosis+and+tubular+atrophy:+what+is+in+a+name.">Chronic allograft nephropathy or interstitial fibrosis and tubular atrophy: what is in a name.</a> Current Opinion in Nephrology and Hypertension. 23 (3), 245-250 (2014).</li><li>Dang, Z., MacKinnon, A., Marson, L. P., Sethi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tubular+atrophy+and+interstitial+fibrosis+after+renal+transplantation+is+dependent+on+galectin-3.">Tubular atrophy and interstitial fibrosis after renal transplantation is dependent on galectin-3.</a> Transplantation. 93 (5), 477-484 (2012).</li><li>Yin, D., 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=Blood+circuit+reconstruction+in+an+abdominal+mouse+heart+transplantation+model.">Blood circuit reconstruction in an abdominal mouse heart transplantation model.</a> Journal of Visualized Experiments. (172), e62007 (2021).</li><li>Zhang, 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=Improved+techniques+for+kidney+transplantation+in+mice.">Improved techniques for kidney transplantation in mice.</a> Microsurgery. 16 (2), 103-109 (1995).</li><li>Mannon, R. B., 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=Chronic+rejection+of+mouse+kidney+allografts.">Chronic rejection of mouse kidney allografts.</a> Kidney International. 55 (5), 1935-1944 (1999).</li><li>Coffman, T., 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=Improved+renal+function+in+mouse+kidney+allografts+lacking+MHC+class+I+antigens.">Improved renal function in mouse kidney allografts lacking MHC class I antigens.</a> Journal of Immunology. 151 (1), 425-435 (1993).</li><li>Martins, P. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Learning+curve,+surgical+results+and+operative+complications+for+kidney+transplantation+in+mice.">Learning curve, surgical results and operative complications for kidney transplantation in mice.</a> Microsurgery. 26 (8), 590-593 (2006).</li></ol>

虽然小鼠的皮肤移植模型对于研究同种免疫排斥事件是简单易行的，但用于更具体地研究心脏16 和肾移植10 后同种免疫相关炎症改变的手术技术已被证明是复杂且非常苛刻的。从移植肾病学家的角度来看，建立一种有效且长期稳定的小鼠肾移植模型对于许多功能和免疫学研究仍然具有不可替代的意义。此外，与其他器官移植相比，小鼠肾移植模型即使存在一定?...

自Sakowitz等人于1973年首次开发肾移植小鼠模型以来1，它已被证明是研究移植缺血性损伤和同种免疫排斥机制以及开发旨在延长同种异体移植存活并可能实现免疫耐受性的新疗法的重要实验工具。然而，手术技术已被证明是复杂且非常苛刻的，有时具有并发症，例如血管吻合口狭窄导致肾前非免疫性肾移植失败2，肾后衰竭引起的缺血和随后移植输尿管坏死，移植输尿管吻合口狭窄和/或受者的尿囊导致尿流出破坏。所有这些都是小鼠肾移植尚未进一步发展的原因，因此未被广泛使用。建立一种有效且长期稳定的小鼠肾移植模型，不伴血管和尿路并发症，对于移植领域的许多研究仍然具有不可替代的意义，重点是肾免疫介导的，但也包括传染病3。此外，与小鼠模型中的其他器官移植如肺、心和肠移植4，5相比，小鼠肾移植模型即使在主要组织相容性抗原差异3，6的情况下也为研究长期生存提供了机会。还表明，在供体 - 受体菌株组合的相同设置中，不同的器官移植（例如心脏或肾脏）具有不同的动力学和同种异体移植物排斥反应的发生3。此外，从肾脏学的角度来看，与简单的皮肤移植实验相比，它是在急性和慢性排斥事件背景下研究实质介导的免疫调节机制的更合适的模型。
根据以前关于小鼠肾移植手术技术的报告3，7，8，9，我们在这里证明了以下可靠的改进，这些改进在过去10年中在我们组内已成功应用10，11，12：首先，输尿管动脉被安全地保存，因为肾动脉被 整体 切除 与腹主动脉的相应部分一起。其次，一种新的，简单而快速的无结血管吻合术技术，其中吻合术的最终缝合线不像传统方法那样与上领带的末端相连，而是保持自由。该技术能够在肾脏再灌注后增加或减少吻合口的大小，以避免血管狭窄和腹腔内出血。第三，使用21 G和30 G注射器针头作为辅助穿刺引导工具，以便将供体输尿管植入受者的膀胱壁，减少对受者膀胱的损伤并促进无狭窄吻合口的形成。
在这份报告中，我们还将传统的，广泛使用的技术与我们实验室建立的改良技术进行了比较，发现肾小管萎缩和肾移植间质组织纤维化的程度没有显着差异。在以前的研究中，我们还将这种新技术与传统方法在局部出血，血栓形成，进行血管吻合术的时间和存活率方面的结果进行了比较。我们发现改善，例如局部血栓形成事件的显着减少（1.1% vs 6.6%），吻合术时间缩短，以及肾脏同源移植物的长期生存率高度可重复（95% vs 84% 使用传统方法）10。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>30G-needles</td>
			<td>Braun</td>
			<td>456300</td>
			<td>-</td>
		</tr>
		<tr>
			<td>acepromazine</td>
			<td>CP Pharma</td>
			<td>Tranquisol P</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Bepanthen eye ointment</td>
			<td>Haus-Apotheke</td>
			<td>PZN 01578675</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Bonn Micro Forceps</td>
			<td>FST</td>
			<td>11083-07</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Box for insulation and oxygen supply device</td>
			<td>RUSKINN</td>
			<td>INVIV</td>
			<td>-</td>
		</tr>
		<tr>
			<td>C57BL/6J&#160; mice</td>
			<td>Charles River. Germany</td>
			<td>no catalog number</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Carprofen</td>
			<td>Zoetis</td>
			<td>Rimadyl 50 mg/ml</td>
			<td>-</td>
		</tr>
		<tr>
			<td>CATHETER-FEP 26G</td>
			<td>TERUMO</td>
			<td>Surflo-W</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Clip Applicator Forceps Style</td>
			<td>FST</td>
			<td>18057-14</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Curved forceps</td>
			<td>WPI</td>
			<td>14114-G</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Cutasept skin disinfection</td>
			<td>VWR</td>
			<td>BODL980365</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Dehydrator</td>
			<td>DIAPATH</td>
			<td>Donatello</td>
			<td>-</td>
		</tr>
		<tr>
			<td>electrosurgical pen</td>
			<td>Bovie</td>
			<td>CHANGE-A-TIP</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Embedding machine</td>
			<td>Wuhan Junjie Electronics Co., Ltd</td>
			<td>JB-P5</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Sinopharm Group Chemical Reagent Co. LtD</td>
			<td>100092683</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Frozen platform</td>
			<td>Wuhan Junjie Electronics Co., Ltd</td>
			<td>JB-L5</td>
			<td>-</td>
		</tr>
		<tr>
			<td>gauze pads, cotton swabs</td>
			<td>Lohmann-Rauscher</td>
			<td>13353</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Glass slide</td>
			<td>Servicebio</td>
			<td>G6004</td>
			<td>-</td>
		</tr>
		<tr>
			<td>HE dye solution set</td>
			<td>Servicebio</td>
			<td>G1003</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Heating mat</td>
			<td>THERMO MAT PRO 30W</td>
			<td>HTP-30</td>
			<td>-</td>
		</tr>
		<tr>
			<td>hemostatic sponge</td>
			<td>CuraSpon</td>
			<td>J1276A</td>
			<td>-</td>
		</tr>
		<tr>
			<td>heparine-solution</td>
			<td>Haus-Apotheke</td>
			<td>PZN 03029820</td>
			<td>-</td>
		</tr>
		<tr>
			<td>ice box</td>
			<td>PETZ</td>
			<td>No Catalog Number available</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Imaging system</td>
			<td>Nikon</td>
			<td>Nikon DS-U3</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Inhalation anesthesia device</td>
			<td>GROPPLER</td>
			<td>BKGM 0616</td>
			<td>-</td>
		</tr>
		<tr>
			<td>isoflurane</td>
			<td>CP Pharma</td>
			<td>Isofluran CP 1 ml/ml</td>
			<td>-</td>
		</tr>
		<tr>
			<td>ketamine</td>
			<td>Zoetis</td>
			<td>no catalog numer</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Masson dye solution set</td>
			<td>Servicebio</td>
			<td>G1006</td>
			<td>-</td>
		</tr>
		<tr>
			<td>metamizole</td>
			<td>WDT</td>
			<td>no catalog numer</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Micro scissors</td>
			<td>FST</td>
			<td>15000-00,15000-10</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Micro Serrefine ( Clamp ) Angled / 16 mm</td>
			<td>FST</td>
			<td>18055-06</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Microscope</td>
			<td>Leica</td>
			<td>LEICAMZ6</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Microscope light</td>
			<td>SCHOTT</td>
			<td>KL2500LED</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Neutral gum</td>
			<td>SCRC</td>
			<td>10004160</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Oven</td>
			<td>Tianjin Laibo Rui Instrument Equipment Co., Ltd</td>
			<td>GFL-230</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Pathology slicer</td>
			<td>Shanghai Leica Instrument Co., Ltd</td>
			<td>RM2016</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Saline solution (NaCl 0.9 %)</td>
			<td>Haus-Apotheke</td>
			<td>PZN 06178437</td>
			<td>-</td>
		</tr>
		<tr>
			<td>scissors</td>
			<td>Peha Instruments</td>
			<td>991083/4</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Slides</td>
			<td>Servicebio</td>
			<td></td>
			<td>-</td>
		</tr>
		<tr>
			<td>small Petri dish</td>
			<td>Sarstedt</td>
			<td>8,33,900</td>
			<td>-</td>
		</tr>
		<tr>
			<td>straight forceps</td>
			<td>WPI</td>
			<td>14113-G</td>
			<td>-</td>
		</tr>
		<tr>
			<td>surgical tape</td>
			<td>BSN</td>
			<td>4120</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Suture Tying Forceps - 10 cm</td>
			<td>FST</td>
			<td>18025-10</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Sutures(10-0)</td>
			<td>Medtronic</td>
			<td>N2540</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Sutures(4-0)</td>
			<td>ETHILON</td>
			<td>V4940H</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Sutures(7-0)</td>
			<td>ETHILON</td>
			<td>1647H</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Syringe (0,3 mL)</td>
			<td>BD</td>
			<td>324826</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Syringe (1 mL)</td>
			<td>BD</td>
			<td>320801</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Tissue spreader</td>
			<td>Zhejiang Kehua Instrument Co., Ltd</td>
			<td>KD-P</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Upright optical microscope</td>
			<td>Nikon</td>
			<td>Nikon Eclipse E100</td>
			<td>-</td>
		</tr>
		<tr>
			<td>xylazine</td>
			<td>Bayer</td>
			<td>Rompun</td>
			<td>-</td>
		</tr>
		<tr>
			<td>Xylene</td>
			<td>Sinopharm Group Chemical Reagent Co. LtD</td>
			<td>10023418</td>
			<td>-</td>
		</tr>
	</tbody>
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a modified surgical technique for kidney transplantation in mice

小鼠肾移植是一种复杂且具有挑战性的手术过程。很少有出版物演示这一行动的关键步骤。因此，本文介绍了该技术并指出了与此操作相关的手术注意事项。此外，还演示了与传统程序相比的重要修改。首先，切除并制备腹主动脉的贴片，以便将包括输尿管动脉在内的肾动脉的近端分叉与供体肾脏整体一起横切 。 这降低了输尿管坏死的风险，并避免了尿路阻塞的发展。其次，展示了一种新的血管吻合术方法，该方法允许操作员在肾移植再灌注已经开始后灵活地增加或减小吻合口的大小。这避免了血管狭窄和腹腔内出血的发展。第三，显示了一种能够吻合脆弱的供体输尿管和受体膀胱的技术，不会引起创伤。采用该方案可以缩短手术时间并减少对受体膀胱的损伤，从而显着提高受体小鼠的手术成功率。

移植后四周，与天然受体对侧肾脏相比，改良技术和常规技术均显示肾小管萎缩的中度体征14，15 （图1）。肾小管萎缩的程度显示两种不同技术之间没有显着差异。与正常的非移植肾脏相比，Masson Goldner的三色染色14，15 的肾脏移植后12周均匀地显示出间质组织纤维化的明显迹象（<strong c...

Watch this Scientific Journal Video about A Modified Surgical Technique for Kidney Transplantation in Mice at JoVE.com

A Modified Surgical Technique for Kidney Transplantation in Mice

该方案提出了一种新的小鼠肾移植手术技术，其重点是改进的动脉吻合术策略。还介绍了一种血管缝合技术，包括一种简单且更安全的输尿管-膀胱吻合术方法。这些修饰缩短了手术时间，提高了小鼠肾移植手术的成功率。

一种用于小鼠肾移植的改良手术技术

Kidney transplantation in mice is a complicated and challenging surgery procedure. There are very few publications demonstrating the key steps of this ...

小鼠肾移植对于研究急性和慢性同种异体移植物排斥反应的机制至关重要。在这里，我们介绍一种新的手术技术，用于小鼠腹部肾移植模型中的血管吻合术和输尿管植入术。在传统的小鼠肾移植模型中，主动脉的上部与肾动脉近似连接。供体的左肾静脉在腔静脉处横切，供体主动脉在肾动脉下方平分，以便稍后吻合受者的腹主动脉。在这种改进的小鼠肾移植技术中，供体左肾静脉也在腔静脉处横切，但供体主动脉在肾动脉的后代上方被切除，并且其吻合口到受体腹主动脉。当主动脉的供体末端结扎时，输尿管动脉被故意保留。现在我们开始演示外科手术。对于麻醉，将小鼠放入盒子中吸入异氟醚约40至60秒，以诱导意识不清。一旦鼠标被麻醉，称量鼠标。根据小鼠体重，腹腔注射氯胺酮，甲苯噻嗪，乙酰丙嗪给予麻醉小鼠。当麻醉效果时，夹住腹部皮毛。然后，将鼠标固定并消毒在手术台上。使用交叉腹部切口切开皮肤。切开腹壁的肌肉。接下来，用盐水吸入纱布小心地移开内脏。暴露左肾、主动脉和下腔静脉。仔细烧灼左腰静脉，包括其下部分支和其他小血管（包括左肾上腺血管）。调动左右肾动脉之间的主动脉区域，长度约为两毫米。分离红外线下腔静脉和主动脉。并在主动脉下方通过弯曲的镊子，在该血管周围放置一个松散的7.06缝合线。使用两个五毫米微血管夹交叉钳夹右肾动脉下方的主动脉区域和下腔静脉。从腔静脉切开左肾静脉。用一毫升肝素溶液冲洗主动脉。收紧结扎，将主动脉切开到结扎下方以及近端钳位下方。精心准备，使脆弱的输尿管动脉完全保存。通过谨慎地烧灼组织周围的所有血管，完全释放左肾和相关血管。取出肾脏并将其以四度储存在细胞系溶液中。初始操作步骤类似于先前显示的供体小鼠的操作步骤，包括麻醉和消毒。通过正中切口打开腹部，然后用盐水溶液用湿纱布覆盖腹部器官。小心保存红外线主动脉，并确保每个大血管分支都经过烧灼。使用电灼术在肾盆近端的位置仔细解剖左输尿管。然后，切除左肾。暴露下腔静脉和腹主动脉，并将其与周围脂肪组织分离，长度约为四毫米。然后交叉钳夹每个与两个微血管钳夹大致和远端。使用微型针座引导10/0单丝缝合针，该针从近端到远端穿过主动脉壁放置。通过轻柔向上牵引缝合线，同时用细弯曲的剪刀直接在针的下表面下方切割，实现约一毫米的椭圆形动脉切开术。以端到端的方式进行供体-受体主动脉吻合术。缝合吻合口的近端和远端。最后一针使用两个单独的10.0缝合线。绑扎后，将两条长缝合线（包括针头）留在原位。在远端 - 近端方向上用两条均匀间隔的缝线连续缝合吻合口的主动脉壁的左侧。最后一次缝合后，引导缝合线穿过上部缝合线领带上方的血管壁的部分厚度，同时对下部缝合线扎带的短端施加轻柔的牵引力。请注意，在这种新的无结技术中，最后一针没有绑在上领带的短端。将移植的肾脏翻转至正常位置。现在，使用近端到远端的三针以近端到远端的方式连续缝制主动脉吻合口的右壁。请注意，与传统的手术技术相比，最后一条缝合线与附近的远端领带合并。不要将其绑在下缝线的末端，将其切割成大约两到三毫米的自由长度。纵向切割下腔静脉，长度足够约1.5毫米。将此切口定位在略低于其主动脉对应物的位置。使用与前面描述相同的缝合程序进行静脉吻合术，不同之处在于吻合口的每一侧需要四到五针。最后的缝线留作与上述主动脉吻合口相似的长度的自由末端。而新的无结吻合技术，最后一针不绑，而是自由。这缩短了手术时间，并允许外科医生增加或减少吻合口的大小。这种改善降低了血管吻合术的难度和术后并发症。完成两次吻合后，使用干拭子对缝合区域施加轻柔的压力约10至20秒。之后，可以移除两个夹子。使用带有10.0缝合线的直针穿透接受者的尿腔，并将其插入21 G针腔进行指导。现在引导21 G针在上一个针应用的位置缝合一个孔，然后拉出针。缝合修剪过的输尿管末端，并在其进入处再次用这种10.0缝合线穿孔膀胱。将10.0细丝和输尿管通过构造的孔拖入尿囊。将供体的输尿管吻合到接受者的尿囊。在这里，将输尿管的外膜连接到膀胱壁的外膜，并用三到四针进行间歇缝合。拉出牵引缝合线。将肠道放回腹腔。进行两层缝合，首先是腹部肌肉，然后是皮肤以关闭腹部伤口。将移植的小鼠放入氧气和温度受控的腔室中，以便在手术后恢复。术后连续四天给予充分的术后镇痛。在移植麻醉后五天对受体小鼠进行对侧肾切除术。抬起并记录鼠标的状态。我们通过评估移植的等移植肾与天然受体对侧肾的组织学变化来比较传统和改进的移植技术。肾小管和肾间质纤维化的奖杯程度在移植后4周和12周没有发现显着差异。我们之前研究了这种新的无结技术的结果，并在程序互操作和术后并发症的技术方面将其与经典方法进行了比较。这里显示的改良技术与移植内动脉静脉血栓形成的发生率较低有关。进行狭窄的时间明显减少，并且实现了出色的肾脏等移植物，实现了长期生存。小鼠肾移植是一个复杂而尖端的过程。这是一种用于血管吻合术和肾植入术的新型手术技术，致力于提高整体成功率，使其成为研究肾移植后自身免疫反应的可靠工具。

Research

JoVE Journal

Immunology and Infection

4D Multimodality Imaging of Citrobacter rodentium Infections in Mice

4D Multimodality Imaging of Citrobacter rodentium Infections in Mice

4D多模态成像<em&gt;柠檬酸rodentium</em在小鼠感染

This protocol outlines the steps required to  longitudinally monitor a bioluminescent bacterial infection using composite 3D diffuse light imaging ...

科研

免疫与感染

Long Term Chronic Pseudomonas aeruginosa Airway Infection in Mice

Long Term Chronic Pseudomonas aeruginosa Airway Infection in Mice

长期慢性<em&gt;铜绿假单胞菌</em&gt;在呼吸道感染小鼠

A mouse model of chronic airway infection is a key asset in cystic fibrosis (CF) research, although there are a number of concerns regarding the model ...

Transplantation of Tail Skin to Study Allogeneic CD4 T Cell Responses in Mice

尾部皮肤来研究异基因CD4 + T细胞应答的小鼠移植

The study of T cell responses and their consequences during allo-antigen recognition requires a model that enables one to distinguish between donor and ...

In Vitro and In Vivo Assessment of T, B and Myeloid Cells Suppressive Activity and Humoral Responses from Transplant Recipients

In Vitro and In Vivo Assessment of T, B and Myeloid Cells Suppressive Activity and Humoral Responses from Transplant Recipients

体外和活体评估移植受体的 T、B 和髓细胞抑制活性和体液反应

The main concern in transplantation is to achieve specific tolerance through induction of regulatory cells. The understanding of tolerance mechanisms ...

Induction of Paralysis and Visual System Injury in Mice by T Cells Specific for Neuromyelitis Optica Autoantigen Aquaporin-4

诱导麻痹及视觉系统损伤小鼠 T 细胞特异的视神经脊髓炎抗原水通道蛋白-4

While it is recognized that aquaporin-4 (AQP4)-specific T cells and antibodies participate in the pathogenesis of neuromyelitis optica (NMO), a human ...

Measuring Deformability and Red Cell Heterogeneity in Blood by Ektacytometry

Ektacytometry 血中红细胞异质性的测定

Decreased red cell deformability is characteristic of several disorders. In some cases, the extent of defective deformability can predict severity of ...

Printed Glycan Array: A Sensitive Technique for the Analysis of the Repertoire of Circulating Anti-carbohydrate Antibodies in Small Animals

印制糖类阵列是分析小动物循环抗碳水化合物抗体汇辑的一种敏感技术

The repertoire of circulating anti-carbohydrate antibodies of a given individual is often associated with its immunological status. Not only the ...

Renal Subcapsular Transplantation of 2'-Deoxyguanosine-Treated Murine Embryonic Thymus in Nude Mice

2'-脱氧核苷在裸体小鼠中治疗鼠胚胎胸腺的肾下囊移植

The thymus is an important central immune organ, which plays an essential role in the development and differentiation of T cells. Thymus transplantation ...

An Immunological Model for Heterotopic Heart and Cardiac Muscle Cell Transplantation in Rats

大鼠异位心脏和心肌细胞移植的免疫模型

Heterotopic heart transplantation in rats has been a commonly used model for diverse immunological studies for more than 50 years. Several modifications ...

Optimization of the Cuff Technique for Murine Heart Transplantation

穆林心脏移植袖口技术的优化

Murine cardiac transplantation has been performed for more than 40 years. With advancements in microsurgery, certain new techniques have been used to ...