Name: a modified cuff technique for mouse cervical heterotopic heart transplantation model
Uploaded: 2022-02-07T15:00:00
Description: Watch this Scientific Journal Video about A Modified Cuff Technique for Mouse Cervical Heterotopic Heart Transplantation Model at JoVE.com

这项工作得到了中国国家自然科学基金（81870304）对李军的支持。

所有动物实验均按照国家研究委员会（美国）2011年《实验动物护理和使用指南》第八版进行。涉及动物受试者的程序已获得中国重庆大学肿瘤医院动物护理和使用委员会的批准。从商业来源获得的重20-30g的雄性BALB / c和C57BL / 6小鼠用于同种异体心脏移植研究。C57BL/6小鼠作为供体和同系受体，BALB/c小鼠作为同种异体受体。该协议的原理图如图1所示。 注意：手?...

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(92), e50753 (2014).</li><li>Cui, D., Tan, C., Liu, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+alternative+technique+of+arterial+anastomosis+in+mouse+heart+transplantation.">An alternative technique of arterial anastomosis in mouse heart transplantation.</a> Clinical Transplantation. 32 (6), 13264 (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 (4), 223-228 (2013).</li><li>Fang, J., 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=A+simplified+two-stitch+sleeve+technique+for+arterial+anastomosis+of+cervical+heterotopic+cardiac+transplantation+in+mice.">A simplified two-stitch sleeve technique for arterial anastomosis of cervical heterotopic cardiac transplantation in mice.</a> American Journal of Translational Research. 5 (5), 521-529 (2013).</li><li>Matsuura, A., Abe, T., Yasuura, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simplified+mouse+cervical+heart+transplantation+using+a+cuff+technique.">Simplified mouse cervical heart transplantation using a cuff technique.</a> Transplantation. 51 (4), 896-898 (1991).</li><li>Corry, R. 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小鼠心脏移植模型有助于研究心脏移植后的排斥机制，有助于开发独特的方法来改善心脏同种异体移植受者的长期生存率。然而，小鼠心脏移植是一项复杂且具有挑战性的任务，需要高水平的显微外科技术，特别是在血管吻合术中11，12，13。小鼠腹部异位心脏移植模型是通过供体主动脉和肺动脉与受体主动脉和下腔静脉吻合缝...

心脏移植已成为终末期心力衰竭的标准治疗方法。每年有超过5，500例心脏移植在国际心肺移植学会注册的组织中进行。在同种异体心脏移植受者中，1 年排斥率仍为 >10%，而 3 年排斥率上升至 36%1，2。然而，对于心脏同种异体移植排斥反应患者缺乏有效的预防性治疗。因此，有必要进行动物模型研究，阐明心脏同种异体移植物免疫和病理排斥反应的生理机制。这些研究将有助于研究开发有效药物所需的新靶点，这将有助于防止心脏同种异体移植排斥反应并提高这些患者群体的存活率。
最近在异位心脏移植的小鼠模型研究中提出了心脏同种异体移植排斥反应的一些潜在的免疫和病理生理机制3，4，5。因此，小鼠异位心脏移植因其与人类基因的高度同源性而成为研究心脏移植后心脏同种异体移植物发生免疫排斥和病理损伤机制的理想临床前模型。普遍的概念是在小鼠模型中使用缝线在受体主动脉和下腔静脉中进行腹部端到端吻合，类似于正常的人体解剖结构。但是，此过程可能会干扰接受者的血液供应和下半身的静脉反流6。因此，本文提出了一种改良的小鼠模型中异位心脏移植程序。
捐赠者的心脏通过使用改进的袖带技术通过端到端颈吻合术植入接受者的颈动脉和颈静脉。这种改良的程序促进了手术的可行性，并确保了心脏移植物的存活，而不会干扰下半身的血液供应和静脉反流。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>&#160;5-0 Polyamide Monofilament suture</td>
			<td>B.Braun Medical Inc.</td>
			<td>C3090954</td>
			<td></td>
		</tr>
		<tr>
			<td>&#160;8-0 Polyamide Monofilament suture</td>
			<td>B.Braun Medical Inc.</td>
			<td>C2090880</td>
			<td></td>
		</tr>
		<tr>
			<td>10-0 Polyamide Monofilament suture</td>
			<td>B.Braun Medical Inc.</td>
			<td>G0090781</td>
			<td></td>
		</tr>
		<tr>
			<td>22 G polyurethane cuff</td>
			<td>B.Braun Medical Inc.</td>
			<td>4251628-02</td>
			<td></td>
		</tr>
		<tr>
			<td>26 G polyurethane cuff</td>
			<td>Suzhou Linhua Medical Instrument Co., LTD</td>
			<td>REF383713</td>
			<td></td>
		</tr>
		<tr>
			<td>Anesthesia induction chamber</td>
			<td>RWD Life Science Co., LTD</td>
			<td>V100</td>
			<td></td>
		</tr>
		<tr>
			<td>Atraumatic microvascular clamp</td>
			<td>Beyotime</td>
			<td>FS500</td>
			<td></td>
		</tr>
		<tr>
			<td>BALB/c and C57BL/6 mice (20&#8211;30 g)</td>
			<td>Centre of Experimental Animals (Army Medical University, Chongqing, China)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>US Biological life Sciences</td>
			<td>352004</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrocoagulator</td>
			<td>Guangzhou Runman Medical Instrument Co., LTD</td>
			<td>ZJ1099</td>
			<td></td>
		</tr>
		<tr>
			<td>Gauze</td>
			<td>Henan piaoan group Co., LTD</td>
			<td>10210402</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Guangzhou Dewei Biological Technology Co., LTD</td>
			<td>DK0032</td>
			<td></td>
		</tr>
		<tr>
			<td>Heparin</td>
			<td>North China Pharmaceutical Co., LTD</td>
			<td>2101131-2</td>
			<td></td>
		</tr>
		<tr>
			<td>HTK solution</td>
			<td>Shenzhen Changyi Pharmaceutical Co., LTD</td>
			<td>YZB/Min8263-2013</td>
			<td></td>
		</tr>
		<tr>
			<td>Injection syringe (10 mL)</td>
			<td>Shandong weigao group medical polymer Co., LTD</td>
			<td>20211001</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>RWD Life Science Co., LTD</td>
			<td>21070201</td>
			<td></td>
		</tr>
		<tr>
			<td>Physiological saline</td>
			<td>Southwest pharmaceutical Co., LTD</td>
			<td>H50021610</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalp needle</td>
			<td>Hongyu Medical Group</td>
			<td>20183150210</td>
			<td></td>
		</tr>
		<tr>
			<td>Shaver</td>
			<td>Beyotime</td>
			<td>FS600</td>
			<td></td>
		</tr>
		<tr>
			<td>Small animal anesthesia machine</td>
			<td>RWD Life Science Co., LTD</td>
			<td>R500</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical operation microscope</td>
			<td>Tiannuoxiang Scientific Instrument Co. , Ltd, Beijing, China</td>
			<td>SZX-6745</td>
			<td></td>
		</tr>
		<tr>
			<td>Swab</td>
			<td>Yubei Medical Materials Co., LTD</td>
			<td>21080274</td>
			<td></td>
		</tr>
	</tbody>
</table>

a modified cuff technique for mouse cervical heterotopic heart transplantation model

心脏同种异体移植排斥反应限制了心脏移植后患者的长期生存。小鼠心脏移植模型是临床前研究中研究心脏同种异体移植排斥反应机制的理想选择，因为它们与人类基因具有高度同源性。这种理解将有助于开发独特的方法来改善用心脏同种异体移植物治疗的患者的长期生存率。在小鼠模型中，腹部供体心脏植入通常使用缝线对受体的主动脉和下腔静脉进行端到端吻合。在该模型中，通过改良袖带技术通过端到端吻合将供体的心脏植入受体的颈动脉和颈静脉。移植手术无需缝合即可进行，因此可以增加接受者的生存，因为不会干扰下半身的血液供应和静脉反流。该小鼠模型将有助于研究心脏同种异体移植物的免疫和病理（急性/慢性）排斥反应的机制。

在该小鼠宫颈异位心脏移植模型中，受体小鼠的存活率约为95.2%（21只小鼠中有20只存活）。死亡的主要原因是术后出血。具有规律节律的快速心跳可作为植入供体心脏存活的指标。
C57BL/6和BALB/c小鼠在该模型中分别为MHC（H-2b）和MHC（H-2d）类型，分别为7，8。这两种菌株因H-2而异，H-2导致急性T细胞介导的排斥反应9?...

Watch this Scientific Journal Video about A Modified Cuff Technique for Mouse Cervical Heterotopic Heart Transplantation Model at JoVE.com

A Modified Cuff Technique for Mouse Cervical Heterotopic Heart Transplantation Model

在本协议中，使用小鼠心脏移植模型来研究心脏同种异体移植排斥的机制。在这种异位心脏移植模型中，提高了手术效率，并使用改进的袖带技术对心脏植入的颈椎端到端吻合确保了心脏移植物的存活。

一种改进的袖带技术用于小鼠颈椎异位心脏移植模型

Cardiac allograft rejection limits the long-term survival of patients after heart transplantation. A mouse heart transplantation model is ideal for ...

小鼠心脏移植模型有助于研究心脏同种异体移植物的免疫学和矢状旁排斥机制。这种理解将有助于开发一种独特的方法来改善患者的长期生存率。这种宫颈移植手术是使用袖带方法在没有缝合的情况下进行的。因此，它降低了血管吻合的复杂性，避免了对下半身血液供应和静脉反流的干扰。演示该程序的将是来自我们实验室的外科医生Xin Mao博士。首先，将受体鼠标放在手术台的加热垫上，并在手术区域剃掉头发。对手术区域进行消毒后，从右下颌角到尾端，做一个平行于颈椎中线的1.5至2厘米切口。使用电凝器和微钳解剖约1厘米的右颈外静脉。用无神经微血管钳夹住近端的静脉，并在远端结扎。将静脉的远端穿过带有斜端和浅凹槽的 22 号聚氨酯倒钩袖带。使用微血管夹用袖带的手柄固定静脉。删除 8-0在远端进行结扎缝合，将内腔翻过由浅表倒钩住的袖带，并用10-0手术缝合线将其固定在表面的凹槽中。切除右舌下腺，形成用于植入心脏移植物的窝，并保留上颌下腺的右叶和右胸锁乳突。使用微钳解剖右颈总动脉约 1 厘米，并在近端用无创伤微血管钳夹夹住动脉。结扎并切断动脉。将动脉的远端穿过带有斜端和表面凹槽的 26 号聚氨酯倒钩袖带。使用微血管夹用袖带的手柄固定动脉。取下远端的结扎缝合线，将腔内翻在袖带上，并用浅表倒钩和凹槽用 10-0 手术缝合线固定。准备好接受者的血管后，在血管上滴100国际单位每毫升肝素盐水以防止血栓形成。用湿盐水纱布覆盖宫颈切口，以便随后植入。使用电动剃须刀剃掉腹毛。对手术区域进行消毒后，沿耻骨联合到剑突下中线切开腹部，并用牵开器扩大切口区域。使用电凝器和微型镊子解剖腹主动脉和下腔静脉1厘米，并通过下腔静脉注射1毫升生理盐水，每毫升肝素补充250国际单位进行肝素化。切除腹主动脉和下腔静脉。用手术剪刀将胸壁分开，沿两侧腋前线切除胸部，然后以 8-0 的比分结扎上腔静脉手术缝合。在上胸下腔静脉插入头皮针。然后从上麻痹下腔静脉通过头皮针注射补充每毫升肝素100单位的冰冷生理盐水，以丰富供体心脏，直到血色褪色。从主动脉弓向供体心脏再灌注两到三毫升冰冷的组氨酸-色氨酸-酮戊二酸溶液，以保护供体心肌。用5-0手术缝合上静脉和下腔静脉以及肺静脉。在分支之前解剖并切断供体主动脉和肺动脉。然后分割上腔静脉和下腔静脉和肺静脉，以切除供体的心脏。将供体心脏以倒置位置植入受体小鼠的颈椎袋中。将带有外翻的颈静脉袖带拉入供体肺动脉腔，以进行端到端吻合术。使用表面上的凹槽通过 10-0 手术缝合线结扎袖带以固定吻合口。采用类似的程序进行供体主动脉与受体颈动脉的端到端吻合术。松开颈静脉的无创伤微血管钳夹，然后松开颈动脉，重新灌注供体的心脏。固定心脏移植物并正确缝合以防止移植物扭曲。使用5-0聚酰胺单丝缝合线用连续缝合线关闭宫颈切口。使用该小鼠宫颈异位心脏移植模型，受体小鼠的存活率约为95.2%，所有心脏同种异体移植物在移植后8天内丢失。相比之下，所有同基因心脏移植存活超过四周。凹槽和袖带可更好地固定外翻的血管，避免反复操作血管。该手术中用于血管吻合术的改良袖带法也适用于不同器官移植动物模型中的另一种吻合术。

Research

JoVE Journal

Medicine

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

用改进的袖套法小鼠颈部心脏移植模型

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are ...

科研

医学

A Modified Method for Heterotopic Mouse Heart Transplantion

对于异位小鼠心脏移植技术的一种改进方法

Mice are often used as heart transplant donors and recipients in studies of transplant immunology due to the wide range of transgenic mice and reagents ...

Murine Heterotopic Heart Transplant Technique

小鼠异位心脏移植技术

It is now over forty years since this technique was first reported by Corry, Wynn and Russell. Although it took some years for other labs to become ...

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 ...

A Novel Microsurgical Model for Heterotopic, En Bloc Chest Wall, Thymus, and Heart Transplantation in Mice

一种新的显微模型异位，整块胸壁，胸腺，和心脏移植小鼠

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression ...

Mouse Model for Pancreas Transplantation Using a Modified Cuff Technique

应用改良袖套技术的小鼠胰腺移植模型

Mouse models have several advantages in transplantation research, including easy handling, a variety of genetically well-defined strains, and the ...

Murine Cervical Aortic Transplantation Model using a Modified Non-Suture Cuff Technique

使用改性非缝合铐技术的宫颈主动脉移植模型

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. ...