这项工作是由澳大利亚资助计划- 1029205国家健康和医学研究委员会（NHMRC）的支持，由澳大利亚Microsearch基金会，由Myee科德林顿医学研究基金会。

在此之前开始实验，获得规划批准的实验从相关机构的动物护理伦理委员会。在按照你的机构的要求，保持小鼠。下面的协议已获得悉尼大学，皇家阿尔弗雷德王子医院委员会。 在此之前开始手术，所有仪器应彻底清洗，并浸泡在80％酒精消毒。一些机构规定蒸压然而，这可以在长期内损害精细的显微外科器械。尽可能使用一次性无菌设备。 1，麻醉<ol><li>麻醉小鼠异氟醚在密闭容器内，然后将其放置在仰卧的工作板，快速连接麻醉鼻锥。麻醉捏后脚，以确保没有反射试验是否足够。 </li><li>剃的皮肤用手术刀片和消毒用80％的乙醇。另外，在70％异丙醇2％氯己定，可用于皮肤杀菌。在麻醉诱导的异氟醚浓度为3％，但减少到1-1.5％进行维护。作出轻微调整，这个浓度，以维持正常呼吸和心脏速率。维持动物的加热板上的温度。 </li></ol> 2，供体手术<ol><li>使开胸通过切割胸部通过胸廓两侧从肋缘至腋下随后在剑突的水平横向切割，以形成胸壁皮瓣，露出心脏和血管。提起皮瓣头来旁边钉选到操作板。撕下心包，暴露心脏和血管。 </li><li>提起下腔静脉钳，一手通过下腔静脉其它注射1ml冷肝素生理盐水近端到心脏，然后把一小动脉夹上腔静脉，以防止措施的灌注液反向通过针孔吨流动。 </li><li>用纱布和棉签，收回心脏向下，显露升主动脉和肺动脉。通过一对microscissors中的一个叶片通过通道（横窦）后到主动脉和肺动脉束并切断主动脉和肺动脉一起尽可能远侧尽可能地确保足够的长度用于吻合术。 </li><li>领带和划分下腔静脉，右侧上腔静脉（SVC），用6-0丝线离开SVC和肺静脉。另配合下腔静脉和右SVC然后将一个线程后到心脏与肺静脉环绕左SVC在一起，配合它。通过切割血管远端的关系，然后将其在冷的无菌盐水中储存于4℃直至移植的供体部位收获心脏。这会导致供体放血死亡。 </li></ol> 3，受体手术<ol><li>麻醉收件人如上面（第1节）。仔细剃腹部，以避免刺激，然后做一个通过剖腹手术从耻骨正中切口，以剑突和收回使用回形针弯曲形成拉钩。包裹肠在温暖的无菌生理盐水浸泡的纱布和缩回到腹部的右上方。 </li><li>揭露基础设施肾主动脉和下腔静脉，主动脉自由和下腔静脉从左侧肾动脉和静脉捆绑段至髂分叉使用烧灼设备从他们的腰椎血管分裂。注意使烧灼是在适当的温度和所用足够的时间来划分和密封该容器端部。 </li><li>应用无创小血管夹到主动脉和下腔静脉段远端和近端。由第一穿刺用30号针头的孔做一个切口在主动脉的前壁;然后切一个切口垂直microscissors相匹配的供体主动脉的大小。冲洗主动脉管腔与肝素生理盐水，以消除任何BLOOD血栓。 </li><li>在供体心脏带给受区覆盖着纱布浸泡在冷盐水，并将其放置到右侧腹部。确保供体主动脉定位在接受​​者主动脉与供体肺动脉位于旁边的收件人IVC旁切口。 </li><li>供体主动脉端 - 侧吻合首先使用运行10-0尼龙缝线从近侧转角和缝合沿左侧开始的收件人主动脉直到到达远端的角落，在该阶段的动物是通过180°的旋转。轻轻的供体心脏移动到腹部的左侧以暴露右侧的主动脉，并通过右侧从远端到近端主动脉壁的继续缝合。收盘前主动脉吻合，轻轻冲洗管腔与肝素生理盐水，以消除任何血块和空气。 </li><li>吻合供体肺动脉（PA）的端至端与接收方腔静脉。做一个切口版tically与下腔静脉在一个站点根据动脉吻合前壁microscissors。吻合的捐赠者PA到收件人IVC与运行从下腔静脉管腔内左侧壁的远端开始10-0尼龙缝线。在到达近端后，继续沿前右侧壁到前端的缝线来完成吻合术。在关闭吻合术，轻轻冲洗管腔以除去任何凝块和空气。 </li><li>释放前用棉签血管钳，地方明胶海绵块周围的吻合部位，并轻轻向下按压直到止血的实现。在再血管化的时候，先松开远侧钳，随后近侧钳。 </li><li>血运重建后，在37℃适用温生理盐水的移植外，协助其恢复。接枝通常会立即开始颤动，并自发地恢复为窦性心律在几分钟之内。注入0.6毫升温暖皮下注射生理盐水，以维持收件人的水合作用。手术前完成注入丁丙诺啡皮下镇痛</li><li>关闭腹部伤口用一个5-0可吸收缝线运行两个图层。首先，完成内层和沿皮肤继续。 </li></ol> 4，恢复和移植物监测<ol><li>注射氨苄青霉素感染的预防和放置在一个加热垫收件人在37℃进行恢复。大多数动物迅速恢复，通常喝，往往在3个小时吃。如果鼠标显示窘迫征象，仔细检查，以确定病因。如果没有明显的原因，治疗与丁丙诺啡和密切监测。请教兽医，如果症状严重或持续时间超过8小时，并考虑安乐死。如果有必要，继续给予丁丙诺啡12小时注射，直到症状消失。如果鼠标显示困难的症状8小时后，给予持续12丁丙诺啡每小时注射，直到他们解决。咨询兽医，如果症状持续时间超过48小时，如果需要安乐死。 </li><li>监测移植心脏的跳动直接腹部触诊，记录节拍的强度+ + + +为​​一个健康的移植到+为弱拍，由于先进的排斥反应及 - 由于完成的排斥移植的不跳动。每日监测小鼠在第10天，然后3次，每周为实验的持续时间。 </li></ol>

<ol>
	<li>Aramaki, O., 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=Interleukin-10+but+not+transforming+growth+factor-beta+is+essential+for+generation+and+suppressor+function+of+regulatory+cells+induced+by+intratracheal+delivery+of+alloantigen.">Interleukin-10 but not transforming growth factor-beta is essential for generation and suppressor function of regulatory cells induced by intratracheal delivery of alloantigen.</a> Transplantation. 79, 568-576 (2005).</li><li>Chen, R. H., Bushell, A., Fuggle, S. V., Wood, K. J., Morris, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+granzyme+A+and+perforin+in+mouse+heart+transplants+immunosuppressed+with+donor-specific+transfusion+and+anti-CD4+monoclonal+antibody.">Expression of granzyme A and perforin in mouse heart transplants immunosuppressed with donor-specific transfusion and anti-CD4 monoclonal antibody.</a> Transplantation. 61, 625-629 (1996).</li><li>Poulin, L. F., 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=Interleukin-9+promotes+eosinophilic+rejection+of+mouse+heart+allografts.">Interleukin-9 promotes eosinophilic rejection of mouse heart allografts.</a> Transplantation. 76, 572-577 (2003).</li><li>Ono, K., Lindsey, E. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+technique+of+heart+transplantation+in+rats.">Improved technique of heart transplantation in rats.</a> J. Thorac. Cardiovasc. Surg. 57, 225-229 (1969).</li><li>Corry, R. J., 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=Primarily+vascularized+allografts+of+heart+in+mice.">Primarily vascularized allografts of heart in mice.</a> Transplantation. 16, 343-350 (1973).</li><li>Larsen, 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=CD40-gp39+interactions+play+a+critical+role+during+allograft+rejection.+Suppression+of+allograft+rejection+by+blockade+of+the+CD40-gp39+pathway.">CD40-gp39 interactions play a critical role during allograft rejection. Suppression of allograft rejection by blockade of the CD40-gp39 pathway.</a> Transplantation. 61, 4-9 (1996).</li><li>Saitovitch, D., Bushell, A., Mabbs, D. W., Morris, P. J., Wood, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinetics+of+induction+of+transplantation+tolerance+with+a+nondepleting+anti-Cd4+monoclonal+antibody+and+donor-specific+transfusion+before+transplantation.+A+critical+period+of+time+is+required+for+development+of+immunological+unresponsiveness.">Kinetics of induction of transplantation tolerance with a nondepleting anti-Cd4 monoclonal antibody and donor-specific transfusion before transplantation. A critical period of time is required for development of immunological unresponsiveness.</a> Transplantation. 61, 1642-1647 (1996).</li><li>Callaghan, C. 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=Regulation+of+allograft+survival+by+inhibitory+FcgammaRIIb+signaling.">Regulation of allograft survival by inhibitory FcgammaRIIb signaling.</a> J. Immunol. 189, 5694-5702 (2012).</li><li>Qian, 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=Impact+of+donor+MHC+Class+I+or+Class+II+antigen+deficiency+on+first-+and+second-set+rejection+of+mouse+heart+or+liver+allografts.">Impact of donor MHC Class I or Class II antigen deficiency on first- and second-set rejection of mouse heart or liver allografts.</a> Immunology. 88, 124-129 (1996).</li><li>Wang, 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=Spontaneous+acceptance+of+mouse+kidney+allografts+is+associated+with+increased+Foxp3+expression+and+differences+in+the+B+and+T+cell+compartments.">Spontaneous acceptance of mouse kidney allografts is associated with increased Foxp3 expression and differences in the B and T cell compartments.</a> Transpl. Immunol. 24, 149-156 (2011).</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> J. Vis. Exp. , (2007).</li><li>Mottram, P. L., 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=Electrocardiographic+monitoring+of+cardiac+transplants+in+mice.">Electrocardiographic monitoring of cardiac transplants in mice.</a> Cardiovasc. Res. 22, 315-321 (1988).</li></ol>

作者什么都没有透露。

小鼠心脏移植是需要相当大的手术技巧掌握一个具有挑战性的显微外科方法。最具挑战性的方面是小直径的血管。此外，有必要限制接收者操作时间和出血。该技术对小鼠心脏移植首先由Corry 等人所述。在1973年，后经莫特拉姆等人 12。我们的修改包括以下几点。首先，供体心脏冷肝素盐水下列开胸主动脉和肺动脉束灌注后不久和横断立即灌注防止热缺血期间收获。其次，从收件人的准备，而不是捐助前准备收件人的捐助收割完成后不久，减少了受援国与生存随之改善腹部暴露时间。最后，使用小型无创伤血管钳到b锁血流量，而不是关系，对收件人主动脉/ IVC束，通常会导致对血管损伤少，并能降低的后肢瘫痪的发病率。 此外，以下几点是成功的重要因素。捐助者和受助者的最佳尺​​寸大约是23〜26 G和最佳的年龄为10至12周。较旧的动物没有较大的血管和他们增加脂肪，使主动脉和下腔静脉的曝光更加困难。必须避免脱水手术过程中的收件人和体温过低。收件人排便需要很好地脱水用温盐水浸泡纱布保护和收件人可以保持在一个加热垫，在37°C。纱布需要在操作期间保持湿润的新鲜，温盐水正规的应用程序。为了尽量减少血液凝块和空气栓塞，可导致后肢麻痹，血管吻合部位必须充分冲洗，关​​闭之前。肠应仔细替换在配备件在移植以防止肠系膜任何扭曲。皮下注射0.6毫升温生理盐水帮助动物恢复，而手术后不久成交量较大静脉注射液可能引起高血压吻合口出血。限制收件人主动脉和下腔静脉交叉钳位至不到30分钟的时间将提高成功率。

器官移植的动物模型可以提供有价值的信息，以提高治疗的临床移植的患者。小鼠模型是研究器官移植排斥或接受免疫机制，由于大范围的转基因小鼠和具体的小鼠不适用于其他动物模型1-3试剂特别有用。与移植小鼠模型的一个挑战是捐助者和受助人的需要相当的技术技能，以达到满意的结果的小尺寸。 一种技术是在大鼠4其后由科里等 5适用于小鼠心脏移植首先描述了心中的异位移植。这项技术涉及的准备之前，供体手术接受者和无灌注供体心脏，这两者都可能损害心脏移植或t的生存他收件人。该过程已被广泛地用作最初描述检查移植排斥和耐受6-8的机制。其他人已经适应了原有的大鼠心脏移植Ono和林赛的心脏移植在小鼠9,10过程。最近，一种技术被发布给小鼠心脏移植是克服了与Corry 等人 11的方法相关联的问题。在这里描述的协议采用了修改，基于莫特拉姆等人 12的方法，该方法包括: 在原位灌注冷肝素生理盐水开胸后立即和事先执行操作供体到受体手术，以减少接受者操作时间。此外，我们使用非创伤性小血管夹，而不是6-0真丝领带。虽然血管夹有采取更多的空间的缺点，他们更容易比的关系来控制，这不应该是太紧或太松并且是那么简单，除去比夹。我们的方法是使用一个单一的连续缝合为容器吻合，虽然在最初的学习阶段的替代方法是使用缝线留在近侧和远侧角，以确保缝合线的均匀性，因此该吻合的通畅。

<table border="0" cellpadding="0" cellspacing="0" width="831">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:293px;">Operating microscope&#160;</td>
			<td style="width:247px;">Leica, Heerbrugg, Switzerland</td>
			<td style="width:124px;">M651</td>
			<td style="width:167px;">10x - 25x magnification</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Anesthetic machine&#160;</td>
			<td>Vet Quip Pty Ltd, Sydney, Australia</td>
			<td style="width:124px;">Vett3</td>
			<td>Capable of delivering a mixture of isoflurane and oxygen in air&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:293px;">Operating board&#160;</td>
			<td style="width:247px;">Hardware store or office supplier</td>
			<td style="width:124px;"></td>
			<td>Dense cork or synthetic capable of taking pins</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:293px;">Heparinised saline (5 U/ml, 4oC)</td>
			<td>Pfizer, USA</td>
			<td style="width:124px;">FW25</td>
			<td>In 1ml syringe with 23 gauge needle on ice</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:293px;">Normal saline (37oC)</td>
			<td style="width:247px;">AstraZeneca Pty Ltd, Australia</td>
			<td style="width:124px;">4538</td>
			<td>In 1ml syringe with 23 gauge needle on warm pad</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:293px;">Sutures 10- nylon, 5-0 Vicryl</td>
			<td>Ethicon, Inc. NJ, USA</td>
			<td style="width:124px;">2870G/J421H</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Buprenorphine (0.05mg/kg in 0.1ml saline)</td>
			<td>Reckitt Benckiser, Sydney, Australia</td>
			<td style="width:124px;"></td>
			<td>In 1ml syringe with 23 gauge needle on ice</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ampcillin (5mg/kg in 0.1 ml saline)&#160;</td>
			<td>Aspen, Sydney, Australia</td>
			<td style="width:124px;"></td>
			<td>In 1ml syringe with 23 gauge needle on ice</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Gel Foam&#160;</td>
			<td>Pharmacia &#38; Upjohn Co. USA</td>
			<td style="width:124px;">801289304</td>
			<td>Cut into small pieces</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">High temperature cautery device&#160;</td>
			<td>Medtronic, USA</td>
			<td style="width:124px;">8444000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Microsurgery instruments:</td>
			<td colspan="2">Shanghai Medical Instruments Co. Ltd.,&#160;&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;&#160;&#160;&#160;&#160;&#160;&#160; micro-needleholders</td>
			<td style="width:247px;">Shanghai, China</td>
			<td style="width:124px;">WT2020</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;&#160;&#160;&#160;&#160;&#160;&#160; micro scissors</td>
			<td>&#160;&#160;&#160;&#160; &#34;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; &#34;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; &#34;</td>
			<td style="width:124px;">WT1020</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:293px;">&#160;&#160;&#160;&#160;&#160;&#160;&#160; micro-forceps (straight tip)</td>
			<td style="width:247px;">&#160;&#160;&#160;&#160; &#34;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; &#34;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; &#34;</td>
			<td style="width:124px;">WA3010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;&#160;&#160;&#160;&#160;&#160;&#160; micro-forceps (curved tip)</td>
			<td>&#160;&#160;&#160;&#160; &#34;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; &#34;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; &#34;</td>
			<td style="width:124px;">WA3020</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;&#160;&#160;&#160;&#160;&#160;&#160; micro-mosquito clamps (1 pair)</td>
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a modified method for heterotopic mouse heart transplantion

老鼠经常被用来作为心脏移植供，受者在移植免疫学研究，由于大范围的转基因小鼠和试剂可用的。的困难，提出由于动物的体积小，参与心脏移植的显微外科的相当大的技术挑战。特别是，移植术后早期高企的技术故障可能导致收件人死亡和术后并发症，如后肢瘫痪或无跳动的心脏。在这里，完整的技术异位小鼠心脏移植证实，涉及收获捐赠的心脏而其随后植入到受体小鼠。供体心脏收获立即就地灌注在与升主动脉和肺动脉的冷肝素生理盐水和横断。接受者操作涉及制备腹主动脉和下腔静脉（IVC），其次是端 - 端吻合术使用一个单一的运行10-0 microsuture和捐助肺动脉与受体下腔静脉的吻合类似的捐助主动脉与受体主动脉。以下操作的动物被注射用0.6 ml生理盐水皮下注射和允许恢复在37℃下加热垫。从227小鼠心脏移植手术的结果总结，成功率在48小时的86.8％。在48小时的13.2％的失败，5（2.2％）出现后肢麻痹，10（4.4％）有因移植缺血性损伤和/或血栓形成非跳动的心脏，15（6.6％），48小时内死亡。

经过最初的培训期，227例小鼠异位心脏移植在我们的小组进行了分析。在最初24小时内，成功率为90.3％，并且在48小时是86.8％。 48小时内的30（13.2％）的故障，5（2.2％）出现后肢瘫痪，不得不被施以安乐死，10（4.4％）有非跳动的心脏，由于移植物缺血损伤和/或血栓形成，而15 （6.6％），48小时内死亡。一些实验接枝残余示于表1的各种应变的组合。同一供体和受体菌株之间的心脏移植已被接受长期而非相同菌株之间移植被拒绝。 移植心脏的外观如图1所示。的心脏移植失败的原因一般是由时间移植后决定的，当心脏停止了跳动，结合心脏的外观。如果需要的话，其原因停止跳动的应该由心脏的组织学分析证实。例如，在一个DBA / 2供体心脏移植的C57BL / 6品系收件人，心脏停止了跳动，第2天，这是最不寻常的排斥反应未致敏受者通常需要至少一个星期。 图1A显示的心脏从这个收件人与血栓形成及心脏组织梗死的证据。在C57BL / 6供体C57BL / 6收件人的应变组合，心脏没有被拒绝，存活&gt; 100天（ 图1B）。在这种情况下，心脏有轻微的大小因肌肉萎缩继发于它的非生命维持状况减少。与此相反，在DBA / 2供体C57BL / 6收件人拒绝的应变组合，移植的心脏已经停止了7天击败并表明拒绝的证据（ 图1C）。这是完全纤维化和萎缩了100天（ 图1D）。 图1:外观血栓，拒绝和无排斥移植的心脏。 （一）DBA / 2心脏移植到C57BL / 6收件人显示移植后的血栓形成和梗死心脏7天。（B）的C57BL / 6心脏移植到C57BL / 6收件人呈现出良好灌注和移植100天后跳动的心脏。（C ）DBA / 2心脏移植到C57BL / 6收件人显示移植后第7天拒绝了心脏。（D）DBA / 2心脏移植到C57BL / 6收件人显示移植100天后拒绝了心脏。箭头表示移植心脏。 <table border="0" cellpadding="0" cellspacing="0" width="489"><tbody><tr><td style="width:138px;">组合</td><td style="width:226px;">存活（天） </td><td style="width:31px;"> Ñ </td><td styLE ="宽度:95px;"&gt; MST（天） </td></tr><tr><td style="width:138px;"> B10.BR→B10.BR </td><td style="width:226px;"> &gt; 200 X7 </td><td style="width:31px;"> 7 </td><td style="width:95px;"> &gt; 200 </td></tr><tr><td style="width:138px;"> C57BL / 6→C​​57BL / 6 </td><td style="width:226px;"> 40，&gt; 200 X2 </td><td style="width:31px;"> 3 </td><td style="width:95px;"> &gt; 200 </td></tr><tr><td style="width:138px;"> F1→B10.BR </td><td style="width:226px;"> 8的2倍，9，12 </td><td style="width:31px;"> 4 </td><td style="width:95px;"> 8.5 </td></tr><tr><td style="width:138px;"> 178.3→B10.BR </td><td style="width:226px;"> 7的2倍，8的2倍，9，10，12，13 </td><td style="width:31px;"> 8 </td><td style="width:95px;"> 8.5 </td></tr><tr><td style="width:138px;"> C57BL / 6→B10.BR </td><td style="width:226px;"> 8×4，9 X2，X5 10，11，12，14×2，15 </td><td style="width:31px;"> 16 </td><td style="width:95px;"> 10 </td></tr><tr><td style="width:138px;">只BALB / c→C57BL / 6 </td><td style="width:226px;"> 7 X9 </td><td style="width:31px;"> 9 </td><td style="width:95px;"> 7 </td></tr><tr><td style="width:138px;"> DBA / 2→C57BL / 6 </td><td style="width:226px;"> 6，7中的10倍</td><td style="width:31px;"> 11 </td><td style="width:95px;"> 7 </td></tr></tbody></table> 表1。心脏移植存活在不同的供体/受体结合。

Watch this Scientific Journal Video about A Modified Method for Heterotopic Mouse Heart Transplantion at JoVE.com

A Modified Method for Heterotopic Mouse Heart Transplantion

A technique is demonstrated for the microsurgical procedure for heterotopic transplantation of hearts in mice, including simplified methods for donor harvesting and recipient vessel anastomosis.

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 ...

a-modified-method-for-heterotopic-mouse-heart-transplantion

Nanyang Technological University

Research

JoVE Journal

Medicine

12.0K 次观看.  University of Sydney. A technique is demonstrated for the microsurgical procedure for heterotopic transplantation of hearts in mice, including simplified methods for donor harvesting and recipient vessel anastomosis.

视频: A Modified Method for Heterotopic Mouse Heart Transplantion

A Modified Heterotopic Swine Hind Limb Transplant Model for Translational Vascularized Composite Allotransplantation (VCA) Research

Vascularized Composite Allotransplantation (VCA) such as hand and face transplants represent a viable treatment option for complex musculoskeletal trauma and devastating tissue loss. Despite favorable and highly encouraging early and intermediate functional outcomes, rejection of the highly immunogenic skin component of a VCA and potential adverse effects of chronic multi-drug immunosuppression continue to hamper widespread clinical application of VCA. Therefore, research in this novel field needs to focus on translational studies related to unique immunologic features of VCA and to develop novel immunomodulatory strategies for immunomodulation and tolerance induction following VCA without the need for long term immunosuppression.
This article describes a reliable and reproducible translational large animal model of VCA that is comprised of an osteomyocutaneous flap in a MHC-defined swine heterotopic hind limb allotransplantation. Briefly, a well-vascularized skin paddle is identified in the anteromedial thigh region using near infrared laser angiography. The underlying muscles, knee joint, distal femur, and proximal tibia are harvested on a femoral vascular pedicle. This allograft can be considered both a VCA and a vascularized bone marrow transplant with its unique immune privileged features. The graft is transplanted to a subcutaneous abdominal pocket in the recipient animal with a skin component exteriorized to the dorsolateral region for immune monitoring.
Three surgical teams work simultaneously in a well-coordinated manner to reduce anesthesia and ischemia times, thereby improving efficiency of this model and reducing potential confounders in experimental protocols. This model serves as the groundwork for future therapeutic strategies aimed at reducing and potentially eliminating the need for chronic multi-drug immunosuppression in VCA.

A Modified Heterotopic Swine Hind Limb Transplant Model for Translational Vascularized Composite Allotransplantation VCA Research

Vascularized Composite Allotransplantations (VCA) have become a clinical reality. However, broad clinical application of VCA is limited by chronic multi-drug immunosuppression. The authors present a reliable and reproducible large animal model to translate novel immunomodulatory strategies that can minimize or potentially eliminate the need of immunosuppression in VCA.

平移血管复合同种异体移植（VCA）研究一种改进的异位猪后肢移植模型

Vascularized Composite Allotransplantation (VCA) such as hand and face transplants represent a viable treatment option for complex musculoskeletal trauma ...

科研

医学

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are available to perform mechanistic in vivo studies. While heart transplantation models using a suture technique were first successfully developed in rats, the translation into an equally widespread used murine equivalent was never achieved due the technical complexity of the microsurgical procedure. In contrast, non-suture cuff techniques, also developed initially in rats, were successfully adapted for use in mice1-3. This technique for revascularization involves two major steps I) everting the recipient vessel over a polyethylene cuff; II) pulling the donor vessel over the formerly everted recipient vessel and holding it in place with a circumferential tie. This ensures a continuity of the endothelial layer, short operating time and very high patency rates4.
Using this technique for vascular anastomosis we performed more than 1,000 cervical heart transplants with an overall success rate of 95%. For arterial inflow the common carotid artery and the proximal aortic arch were anastomosed resulting in a retrograde perfusion of the transplanted heart. For venous drainage the pulmonary artery of the graft was anastomosed with the external jugular vein of the recipient5.
Herein, we provide additional details of this technique to supplement the video.

The murine cervical heart transplantation model is well suited for immunological as well as ischemia reperfusion injury studies. We modified the procedure using a non-suture cuff technique and performed more than 1,000 successful transplants with this approach.
Herein, we provide additional details of this technique to supplement the video.

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 ...

Murine Heterotopic Heart Transplant Technique

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 proficient in and utilize this technique, it is now widely used by many laboratories around the world. A significant refinement to the original technique was developed and reported in 2001 by Niimi. Described here are the techniques that have evolved over more than a decade in the hands of three surgeons (Plenter, Grazia, Pietra) in our center. These techniques are now being passed on to a younger generation of surgeons and researchers.
Based largely on the Niimi experience, the procedures used have evolved in the fine details - details which we will endeavor to relate here in such a way that others may be able to use this very useful model. Like Niimi, we have found that a video aid to learning is a priceless resource for the beginner.

The manuscript describes the steps required to perform the heterotopic heart transplant in the mouse.

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

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed results. However, the cellular and molecular mechanisms of the neotissue development, valve thickening, and stenosis development are not researched extensively. To answer the above questions, we developed a murine heterotopic heart valve transplantation model. A heart valve was harvested from a valve donor mouse and transplanted to a heart donor mouse. The heart with a new valve was transplanted heterotopically to a recipient mouse. The transplanted heart showed its own heartbeat, independent of the recipient&#8217;s heartbeat. The blood flow was quantified using a high frequency ultrasound system with a pulsed wave Doppler. The flow through the implanted pulmonary valve showed forward flow with minimal regurgitation and the peak flow was close to 100 mm/sec. This murine model of heart valve transplantation is highly versatile, so it can be modified and adapted to provide different hemodynamic environments and/or can be used with various transgenic mice to study neotissue development in a tissue engineered heart valve.

In order to understand the cellular and molecular mechanisms underlying neotissue formation and stenosis development in tissue engineered heart valves, a murine model of heterotopic heart valve transplantation was developed. A pulmonary heart valve was transplanted to recipient using the heterotopic heart transplantation technique.

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

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft involves anastomosis of donor inferior vena cava (IVC) to recipient IVC, and donor ascending aorta (Ao) to recipient abdominal Ao. The right upper and middle lung lobes are preserved and function as conduits for blood flow from right heart to left heart.
There are several advantages to using this technique, and it lends itself to a broad range of applications. Because the graft is transplanted in a configuration that allows for dyamic volume-loading, cardiac function may be directly assessed in vivo. The use of pressure-volume conductance catheters permits characterization of load-dependent and load-independent hemodynamic parameters. The graft may be converted to a loaded configuration by applying a clamp to the recipient&#8217;s infra-hepatic IVC. We describe modified surgical techniques for both donor and recipient operations, and an ideal myocardial protection strategy. Depending on the experimental aim, this model may be adapted for use in both acute and chronic studies of graft function, immunologic status, and variable ventricular loading conditions. The conducting airways to the transplanted lung are preserved, and allow for acute lung re-ventilation. This facilitates analysis of the effects of the mixed venous and arterial blood providing coronary perfusion to the graft.
A limitation of this model is its technical complexity. There is a significant learning curve for new operators, who should ideally be mentored in the technique. A surgical training background is advantageous for those wishing to apply this model. Despite its complexity, we aim to present the model in a clear and easily applicable format. Because of the physiologic similarity of this model to orthotopic transplantation, and its broad range of study applications, the effort invested in learning the technique is likely to be worthwhile.

We describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The transplant configuration results in a partially loaded graft circulation, allowing direct functional assessment. This model may be employed for acute or chronic studies of function and immunologic status of the transplanted graft.

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

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&#8217;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.

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.

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

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression must be pursued. Employing vascularized bone marrow transplantation and co-transplantation of the thymus have shown promise in this regard in various animal models.1-11 Vascularized bone marrow transplantation allows for the uninterrupted transfer of donor bone marrow cells within the preserved donor microenvironment, and the incorporation of thymus tissue with vascularized bone marrow transplantation has shown to increase T-cell chimerism ultimately playing a supportive role in the induction of immune regulation. The combination of solid organ and vascularized composite allotransplantation can uniquely combine these strategies in the form of a novel transplant model. Murine models serve as an excellent paradigm to explore the mechanisms of acute and chronic rejection, chimerism, and tolerance induction, thus providing the foundation to propagate superior allograft survival strategies for larger animal models and future clinical application. Herein, we developed a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique. The experience in syngeneic and allogeneic transplant settings is described for future broader immunological investigations via an instructional manuscript and video supplement.

To study combined solid organ and vascularized composite allotransplantation, we describe a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique.

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 ...

A Simplified Model for Heterotopic Heart Valve Transplantation in Rodents

There is an urgent clinical need for heart valve replacements that can grow in children. Heart valve transplantation is proposed as a new type of transplant with the potential to deliver durable heart valves capable of somatic growth with no requirement for anticoagulation. However, the immunobiology of heart valve transplants remains unexplored, highlighting the need for animal models to study this new type of transplant. Previous rat models for heterotopic aortic valve transplantation into the abdominal aorta have been described, though they are technically challenging and costly. For addressing this challenge, a renal subcapsular transplant model was developed in rodents as a practical and more straightforward method for studying heart valve transplant immunobiology. In this model, a single aortic valve leaflet is harvested and inserted into the renal subcapsular space. The kidney is easily accessible, and the transplanted tissue is securely contained in a subcapsular space that is well vascularized and can accommodate a variety of tissue sizes. Furthermore, because a single rat can provide three donor aortic leaflets and a single kidney can provide multiple sites for transplanted tissue, fewer rats are required for a given study. Here, the transplantation technique is described, providing a significant step forward in studying the transplant immunology of heart valve transplantation.

This protocol describes a simple and efficient method for the transplantation of aortic valve leaflets under the renal capsule to allow for the study of alloreactivity of heart valves.

啮齿动物异位心脏瓣膜移植的简化模型

There is an urgent clinical need for heart valve replacements that can grow in children. Heart valve transplantation is proposed as a new type of ...

A Modified Simple Method for Induction of Myocardial Infarction in Mice

Myocardial infarction (MI) represents one of the leading causes of death. MI models are widely used for investigating the pathomechanisms of post-MI remodeling and evaluation of novel therapeutics. Different methods (e.g., isoproterenol treatment, cryoinjury, coronary artery ligation, etc.) have been used to induce MI. Compared with isoproterenol treatment and cryoinjury, coronary artery ligation may better reflect the ischemic response and chronic remodeling after MI. However, traditional methods for coronary ligation in mice are technically challenging. The current study describes a simple and efficient process for induction of MI in mice with readily available materials. The mouse chest skin was cut open under stable anesthesia. The heart was immediately externalized through the intercostal space after blunt separation of the pectoralis major and pectoralis minor. The left anterior descending branch (LAD) was ligated with a 6-0 suture 3 mm from its origin. Following LAD ligation, staining with 2,3,5-Triphenyltetrazolium chloride (TTC) indicated successful induction of MI and temporal changes of post-MI scar size. Meanwhile, survival analysis results showed overt mortality within 7 days after MI, mainly due to cardiac rupture. Moreover, post-MI echocardiographic assessment demonstrated successful induction of contractile dysfunction and ventricular remodeling. Once mastered, an MI model can be established in mice within 2-3 min with readily available materials.

Under adequate anesthesia, the mouse heart was externalized through the intercostal space, and myocardial infarction was successfully induced by ligating the left anterior descending artery (LAD) using materials readily available in most laboratories.

一种改良的小鼠心肌梗死诱导简易方法

Myocardial infarction (MI) represents one of the leading causes of death. MI models are widely used for investigating the pathomechanisms of post-MI ...

A Porcine Heterotopic Heart Transplantation Protocol for Delivery of Therapeutics to a Cardiac Allograft

Cardiac transplantation is the gold standard treatment for end-stage heart failure. However, it remains limited by the number of available donor hearts and complications such as primary graft dysfunction and graft rejection. The recent clinical use of an ex vivo perfusion device in cardiac transplantation introduces a unique opportunity for treating cardiac allografts with therapeutic interventions to improve function and avoid deleterious recipient responses. Establishing a translational, large-animal model for therapeutic delivery to the entire allograft is essential for testing novel therapeutic approaches in cardiac transplantation. The porcine, heterotopic heart transplantation model in the intraabdominal position serves as an excellent model for assessing the effects of novel interventions and the immunopathology of graft rejection. This model additionally offers long-term survival for the pig, given that the graft is not required to maintain the recipient's circulation. The aim of this protocol is to provide a reproducible and robust approach for achieving ex vivo delivery of a therapeutic to the entire cardiac allograft prior to transplantation and provide technical details to perform a survival heterotopic transplant of the ex vivo perfused heart.

We present a protocol for utilizing a normothermic ex vivo sanguinous perfusion system for the delivery of therapeutics to an entire cardiac allograft in a porcine heterotopic heart transplant model.

一种猪异位心脏移植方案，用于向心脏同种异体移植物提供治疗药物

Cardiac transplantation is the gold standard treatment for end-stage heart failure. However, it remains limited by the number of available donor hearts ...