Name: blood circuit reconstruction in an abdominal mouse heart transplantation model
Uploaded: 2021-06-03T13:00:00
Description: Watch this Scientific Journal Video about Blood Circuit Reconstruction in an Abdominal Mouse Heart Transplantation Model at JoVE.com

我们感谢徐云博士作为配音演员彭建华博士对视频编辑的帮助，感谢安妮卡·库克哈恩博士对她的评价和支持。这项工作部分得到了德国研究基金会（DFG）的支持，以促进国际合作（HO2581/4-1对AH）和中国国家科学基金会（NSFC;#81760291到FJ）。

所有动物实验都是按照欧洲议会2010/63/欧盟关于保护用于科学目的的动物的指令（伦理委员会批准，#G1071/09）的指导方针进行的。
注：初步准备、麻醉、术后护理和监测工作与传统手术方法1、2、4相同。BALB/c 小鼠充当心脏捐献者，C57BL/6J 担任移植接受者。小鼠的年龄为8-12周，移植时体重约30克，在标准条?...

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	<li>Corry, R., Winn, H., Russell, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Primarily+vascularized+allografts+of+heart+in+mice.">Primarily vascularized allografts of heart in mice.</a> Transplantation. 16, 343-350 (1973).</li><li>Habertheuer, 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=Donor+tissue-specific+exosome+profiling+enables+noninvasive+monitoring+of+acute+rejection+in+mouse+allogeneic+heart+transplantation.">Donor tissue-specific exosome profiling enables noninvasive monitoring of acute rejection in mouse allogeneic heart transplantation.</a> Journal of Thoracic and Cardiovascular Surgery. 155 (6), 2479-2489 (2018).</li><li>Plenter, R. J., Zamora, M. R., Grazia, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Four+decades+of+vascularized+heterotopic+cardiac+transplantation+in+the+mouse.">Four decades of vascularized heterotopic cardiac transplantation in the mouse.</a> Journal of Investigative Surgery. 26, 223-228 (2013).</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>Alamran, F. G., Shahkolahi, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Total+arterial+anastomosis+heterotopic+heart+transplantation+model.">Total arterial anastomosis heterotopic heart transplantation model.</a> Transplantation Proceedings. 45, 625-629 (2013).</li><li>Melanie, L., et al., Moffatt-Bruce, S. 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=Potential+of+Heterotopic+Cardiac+Transplantation+in+Mice+as+a+Model+for+Elucidating+Mechanisms+of+Graft+Rejection.">Potential of Heterotopic Cardiac Transplantation in Mice as a Model for Elucidating Mechanisms of Graft Rejection.</a> Cardiac Transplantation. , (2012).</li><li>Westhofen, 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=The+heterotopic+heart+transplantation+in+mice+as+a+small+animal+model+to+study+mechanical+unloading+-+Establishment+of+the+procedure,+perioperative+management+and+postoperative+scoring.">The heterotopic heart transplantation in mice as a small animal model to study mechanical unloading - Establishment of the procedure, perioperative management and postoperative scoring.</a> PLoS One. 14 (4), 0214513 (2019).</li><li>Wang, C., Wang, Z., Allen, R., Bishop, G. A., Sharland, A. 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+Modified+Method+for+Heterotopic+Mouse+Heart+Transplantion.">A Modified Method for Heterotopic Mouse Heart Transplantion.</a> Journal of Visualized Experiments. (88), e51423 (2014).</li><li>Mokbel, 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=Histidine-Tryptophan-Ketoglutarate+Solution+for+Donor+Heart+Preservation+Is+Safe+for+Transplantation.">Histidine-Tryptophan-Ketoglutarate Solution for Donor Heart Preservation Is Safe for Transplantation.</a> Annals of Thoracic Surgery. 109 (3), 763-770 (2020).</li><li>Song, 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=Modified+Suture+Technique+in+a+Mouse+Heart+Transplant+Model.">Modified Suture Technique in a Mouse Heart Transplant Model.</a> Asian Journal of Surgery. 34 (2), 86-91 (2011).</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=Assessment+of+graft+function+in+rodent+models+of+heart+transplantation.">Assessment of graft function in rodent models of heart transplantation.</a> Microsurgery. 28 (7), 565-570 (2008).</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=cold+storage+using+a+new+histidine-tryptophan-ketoglutarate-based+preservation+solution+in+isogeneic+cardiac+mouse+grafts.">cold storage using a new histidine-tryptophan-ketoglutarate-based preservation solution in isogeneic cardiac mouse grafts.</a> European Heart Journal. 32 (4), 509-516 (2011).</li><li>T&#252;rk, T. 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=Reduction+of+chronic+graft+injury+with+a+new+HTK-based+preservation+solution+in+a+murine+heart+transplantation+model.">Reduction of chronic graft injury with a new HTK-based preservation solution in a murine heart transplantation model.</a> Cryobiology. 64 (3), 273-278 (2012).</li></ol>

小鼠异位心脏移植手术技术极具挑战性，这涉及到这种模式的广泛应用受到限制。
传统技术的缺点之一是捐赠者肺动脉（PA）的长度有限。它通常长度约为2毫米，而我们模型中使用的供体心脏IIVC的长度一般约为1厘米（图2）。这意味着，在修改后的模型中，IIVC肛门切除术提供了更清晰的视线，使肛门手术得到改善，并防止过于严格的缝合线，甚至损害?...

小鼠异位性腹部心脏移植手术技术是移植免疫学1、2、3研究的标准模型。然而，这是非常具有挑战性的执行，这涉及到限制广泛使用这种模型4，5。
在传统的小鼠心脏移植（THTx）中，供体主动脉和接受者腹部主动脉为肛门，而肺动脉为接受者低维纳卡瓦6、7、8。
在这种修改后小鼠心脏移植技术中，供体主动脉与接受者腹部主动脉分离，供体IIVC与接受者低劣的静脉卡瓦（3、4、6）（图2和图3）有关。

<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>BALB/c AnNCrl mice</td>
			<td>Charles River. Germany</td>
			<td>no catalog number</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>custodiol/HTK</td>
			<td>Dr. Franz K&#246;hler Chemie</td>
			<td>no catalog numer</td>
			<td></td>
		</tr>
		<tr>
			<td>Cutasept skin disinfection</td>
			<td>VWR</td>
			<td>BODL980365</td>
			<td></td>
		</tr>
		<tr>
			<td>electrosurgical pen</td>
			<td>Bovie</td>
			<td>CHANGE-A-TIP</td>
			<td></td>
		</tr>
		<tr>
			<td>gauze pads, cotton swabs</td>
			<td>Lohmann-Rauscher</td>
			<td>13353</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>Inhalation anesthesia device</td>
			<td>GROPPLER</td>
			<td>BKGM 0616</td>
			<td></td>
		</tr>
		<tr>
			<td>insulation and oxygen supply device</td>
			<td>RUSKINN</td>
			<td>INVIV</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>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>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>small Petri dish</td>
			<td>Sarstedt</td>
			<td>833900</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>xylazine</td>
			<td>Bayer</td>
			<td>Rompun</td>
			<td></td>
		</tr>
	</tbody>
</table>

blood circuit reconstruction in an abdominal mouse heart transplantation model

小鼠异位性腹部心脏移植手术技术是移植免疫学研究的标准模型。在这里，介绍了异位性腹部心脏移植模型中经过修饰的血液回路重建的既定技术。这种方法使用胸内低静脉卡瓦（IIVC），而不是供体心脏的肺动脉，用于接受者低等静脉卡瓦的解剖。它促进和提高小鼠腹部心脏移植的成功率。

在这里，介绍了一种经过改进的小鼠异位腹部心脏移植技术，该技术以前在我们的实验室中已经开发出来，并且在过去16年中被证明是有用的。此前，据报道，在总共40例静脉卡瓦到静脉卡瓦（V-V组）相比，40例传统肺动脉静脉卡瓦（P-V组）解剖程序4（表1），船舶解剖病在V-V组服用20.8±1.3分钟， 明显低于P-V组（27.5±1.3分钟，第0.01页<）。暖缺血时间、总接受者...

Watch this Scientific Journal Video about Blood Circuit Reconstruction in an Abdominal Mouse Heart Transplantation Model at JoVE.com

Blood Circuit Reconstruction in an Abdominal Mouse Heart Transplantation Model

演示了异位小鼠心脏移植模型中血液回路重建的新技术。

腹部小鼠心脏移植模型中的血液回路重建

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the ...

小鼠心脏移植是研究异位免疫反应的宝贵模型，引进了小鼠异位心脏移植的新型手术技术。在传统的小鼠心脏移植中，供体主动脉和接受者腹部主动脉为肛门。肺动脉是与接受者低劣的静脉卡瓦的同位素。在这个新的小鼠心脏移植。供体主动脉是接受者腹部主动脉的肛门，捐赠者在胸腔内劣质静脉卡瓦是接受者低等静脉卡瓦的肛门。现在我们开始实验示范。将鼠标放入箱中进行异氟烷吸入和麻醉后，称重鼠标。根据小鼠的体重，给麻醉小鼠注射氯胺酮二甲苯丙胺。麻醉发生时，在手术台上修复和消毒鼠标。将皮肤从颈部切到下腹部。切开腹壁的肌肉，轻轻地将内脏移开。接下来，暴露劣质的维纳卡瓦。将 0.4 毫升肝素溶液缓慢注射到下静脉卡瓦中，等待一分钟后再拔出针头。拔出针头，用剪刀切开劣质静脉卡瓦和腹部主动脉，加速切除。通过执行 U 形切口打开胸腔。完全暴露心脏、肺和所有胸血管。暴露胸腔主动脉。切掉一半的流明。将灌溉管插入胸动脉的开口。切断肺静脉，促进灌溉和排水。注射至少两毫米的四度HTK溶液。心脏完全停止跳动，直到肺静脉流出完全清晰。拔出灌溉管，分离胸骨。取出胸腺。轻轻剥去主动脉拱门周围的脂肪。用10个零缝合线暴露和液合动脉脉冲的躯干。分离事实和连接组织附加在触觉内劣维纳卡瓦暴露和石灰与七零缝合的上级静脉卡瓦，并削减它背后的结条。在主动脉拱门下的心底处做七个零缝合线。劣质静脉卡瓦和两个尿道，然后石灰肺动脉分支和静脉肺血管。尽可能解剖主动脉拱门，但请确保你有一个清晰的流明，没有侧枝的开口。隔膜附近的连体和触内劣质静脉卡瓦下的肺血管。现在，把心脏从胸口取出。将被喷出的捐赠者的心脏放入4度冷硬盘溶液中，并暂时保存。一定不要延长冷缺血时间超过60分钟。现在，我们开始演示受体操作。最初的操作步骤与之前显示的供体小鼠类似，包括麻醉和绝育。腹部皮肤以横向切割的方式。肠道解剖后，用盐水溶液用湿纱布覆盖腹部器官。暴露劣质静脉卡瓦和腹部主动脉，并释放他们从周围的脂肪组织。如有必要，在肾血管下对侧分支容器进行液化或电化。在大脑上部插入右侧的第一个微止血力，在肾脏血管下方插入主动脉。在大脑和主动脉的下部插入右侧的第二个微止血钳。主动脉和静脉空间在两个止血钳之间有超过一厘米的动脉瘤和静脉空间。切入主动脉，比上夹更接近下夹。此外，您可以使用 30G 针做一个小孔，并用微型剪刀打开它。将心脏放入腹腔。主动脉在视觉和静脉卡瓦在另一边，并覆盖它与一个小湿纱布垫。适应和缝合接受者主动脉的捐赠主动脉，开始审计。打个结，然后用一条运行缝合线到这个切口的顶部，大约四到五针。接下来，将心脏向右翻转，再次盖住它，并继续在左侧的缝合，直到达到结尾和结。使用微型剪刀在接受者的腹部劣质静脉卡瓦上切一个圆孔，该洞应具有与捐赠者的体内劣维纳卡瓦流明相同的大小。切口应位于计划主动脉解剖的颅骨。使静脉切口大于主动脉切口。冲洗捐赠者的触内劣质静脉卡瓦。将捐赠者内在性低劣的静脉卡瓦缝制到接受者静脉卡瓦开始。打个结，执行运行缝合，直到切口的顶部达到。使用五针，继续左侧的缝合。最后，在尾角打个结，小心地拧紧。小心不要拉得太紧。将止血海绵的一小部分放在静脉和主动脉瘤周围。先取出下部，然后取出上微止血夹，用温暖的0.9%氯化钠冲洗腹腔。取出止血海绵。观察移植心脏的心跳。如果一切都正确，心脏将再次开始收缩。将肠子放回腹腔，用四个零缝合。为移植的小鼠提供温暖富氧的环境。在以前的研究工作中，我们报告了总共40例静脉卡瓦和静脉卡瓦的比较，以及40例传统肺动脉静脉腔解剖手术。与传统的肺动脉静脉瘤组相比，静脉卡瓦到静脉卡瓦组血管解剖的手术时间要短得多。小鼠异位性腹部心脏移植手术技术是移植免疫学研究的标准模型。这里介绍的这种新颖的血液回路重建技术促进了小鼠的腹部心脏移植。这被称为与减少温暖缺血和重新拍打时间的嫁接心脏，从而提高这种模式的成功率。

Research

JoVE Journal

Medicine

A Mouse Model of in Utero Transplantation

A Mouse Model of in Utero Transplantation

一个小鼠模型在子宫内移植

The transplantation of stem cells and viruses in utero has tremendous potential for treating congenital disorders in the human fetus.  For example, in ...

科研

医学

Orthotopic Liver Transplantation in Rats

大鼠原位肝移植

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

采用实时定量PCR检测供体细胞植入一个具有竞争力的小鼠骨髓移植模型

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation

肺动脉瓣使用异位心脏移植小鼠模型移植

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed ...

Rat Heterotopic Abdominal Heart/Single-lung Transplantation in a Volume-loaded Configuration

大鼠异位心脏腹部/在卷装配置单肺移植

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft ...

Heterotopic Cervical Heart Transplantation in Mice

颈椎异位心脏移植小鼠

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies ...

Apical Resection Mouse Model to Study Early Mammalian Heart Regeneration

根尖切除小鼠模型来研究早期哺乳动物心脏再生

Cardiovascular disease plagues the whole world due to intensive lifestyle changes. Heart regeneration holds great promise for repairing and restoring ...

Using the Sleeve Technique in a Mouse Model of Aortic Transplantation - An Instructional Video

套筒技术在主动脉移植小鼠模型中的应用-教学视频

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the ...

A Chronic Cardiac Ischemia Model in Swine Using an Ameroid Constrictor

应用 Ameroid 的猪慢性心肌缺血模型

Cardiovascular disease remains the number one cause of mortality in the United States. There are numerous approaches to treating these diseases, but ...

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta

球囊损伤诱发小鼠腹主动脉 Myointimal 增生的研究

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine ...