作者没有致谢。

所有动物实验都是遵照德国动物保护法 (TierSchG) 进行的, 并得到当地动物福利委员会 (下萨克森国家消费者保护和食品安全办公室的批准, 协议TSA 14/1556)。适合该模型的小鼠的最小重量为25克. 
 1. 术前准备 注意: 所有的程序都在清洁、消毒的条件下进行, 并带有蒸压仪器. 
<ol> <li> 将 50-60 8 厘米长的丙烯中空纤维平行放置在试管中, 并与两侧的 T 型适配器连接。用胶水填充空心纤维和 T 型适配器的外部分支之间的空间. 
	<ol> <li> 允许至少24小时以使胶水变硬。使用标准切片切断从 T 型适配器延伸出来的中空纤维, 并将硅胶管拉到连接点. </li>
	</ol> </li> <li> 将27克金属针 (从 26G branule) 和 #160 插入一个2法郎的聚氨酯套管。使用显微镜下的手术刀片和微剪刀 (8-12X 放大), 使三至四 fenestrations, 每在远端三分之一的套管, 以确保最佳的静脉回流流量. 
	<ol> <li> 在完成时移除导线。用棉签对结构进行钝夹层和缩回。使用纱布拭子 (5 x 5 cm 2 ) 来吸收多余的液体, 以防止组织脱水. </li>
	</ol> </li> <li> 准备启动卷。为完成, 增加30单位肝素每毫升启动液和2.5% 伏/v 的8.4% 解决方案的 NaHCO 3 的缓冲。将此解决方案存储在4和 #176; C 直到使用. </li>
	<li> 填入 CPB 电路 (如: , 泵, 空气捕手, 油管和两个套管) 与850和 #181; 通过静脉导管启动溶液。一旦填充, 保持体外循环循环, 直到动物准备好插管. </li>
</ol> 2。动物麻醉 <ol> <li> 要进行麻醉, 请将该动物置于2.5% 伏/伏异氟烷/氧混合物的诱导室中。通过评估踏板撤退反射, 尾部捏反射, 和眼睛眨眼反射来确认正确的麻醉。应用兽医眼膏避免眼部干燥. </li>
	<li> 将动物置于温度调节功能的暖垫上。使用插入直肠的探头测量体温, 并连接到数据采集系统. </li>
	<li> 在完全麻醉完成后, 用20G 塑料 braunule 将动物 orotracheally 插管, 将其插入口腔, 并在视觉控制下将其推入气管。使用与鼠标呼吸器连接的异氟醚蒸发器启动机械通气。根据动物的重量, 调整通风, 使潮气量 250-350 和 #181; 我实现了. </li>
	<li> 确保完全镇痛, 注射5毫克/千克动物体重卡洛芬皮下注射。建议将异氟醚浓度保持在 1.3-2.5% 之间的氧。异氟醚浓度可根据手术阶段手动调节. </li>
</ol> 3。外科手术程序 <ol> <li> 在完全麻醉和插管完成后, 用锋利的细剪刀、以钝的方式分离肌肉、并暴露右颈静脉和左颈动脉, 在颈部进行中线皮肤切口。在制备过程中, 使用与双极性钳连接的兽医凝固将小血管凝固, 以确保最小的失血. </li>
	<li> 经颈静脉血管制备后, cranially 结扎左颈总动脉远端段使用8-0 丝缝合线进行分岔. </li>
	<li> 使用硅胶连接器 (0.5 mm ID, 1 mm 外径) 将27克套管的远端连接到动脉流入管上, 在动脉的近段处放置8-0 丝缝线滑动节, 并将套管尖端插入颈动脉. </li>
	<li> 在正确放置套管尖端后, 将套管尖端向前推进 3-4 毫米, 使其下部向主动脉弓。用8-0 丝结固定好套管的尖端. </li>
	<li> 使用显微外科钳和剪刀, 进行钝和尖锐的解剖, 暴露右颈静脉通过钝性准备组织侧向胸锁乳突肌, 接近锁骨, 并放置一个8-0 丝结在远端和一个8-0在近端循环. </li>
	<li> 结扎颈静脉远端后, 将静脉导管尖端放置在右颈静脉, 朝向右心房进展, 并使用丝结固定。为了获得最佳的静脉回流, 可能需要将静脉尖端进入右心室. </li>
	<li> 一旦达到正确的插管位置, 通过直接静脉注射到颈静脉内, 将每克动物体重增加2.5 的肝素, 进行全身肝. </li>
	<li> 用于 real-time 侵入性压力监测, cannulate 左股动脉。在25X 放大倍数下, 暴露腹股沟区, 轻轻地将股股静脉和股神经分开. 
	<ol> <li> 结扎股动脉远端的部分。用微钳在前壁上临时咬合, 用滑结做一个小切口. </li>
		<li> 之后, 在股动脉中插入一根1法郎的聚氨酯套管, 用丝10-0 缝线固定。本套管用于抽取血液样本进行血气分析. </li>
	</ol> </li> <li> 在成功放置动脉和静脉套管后, 通过打开泵启动体外循环。从插管到 CPB 开始的时间大约是45分钟, 开始使用0.5 毫升/分钟的流速, 这取决于系统压力测量, 并增加泵的速度在2分钟内的全流量 (4-6 毫升/分钟) 的血流量. </li>
	<li> 在完全监视下, 使用锐剪刀从柄开始执行上部胸骨, 并在尾部方向上5毫米。立即凝固胸骨边缘以避免流血。通过在颅方向上拉右颈动脉来暴露主动脉弓. </li>
	<li> 为了获得最佳控制, 圆周从胸腺和周围组织中释放主动脉弓, 以便于夹紧。在升主动脉周围放置一个8-0 丝环。要将十字钳放置在心脏停搏液中, 在颅骨方向上拉上丝环, 以更好地暴露升主动脉. </li>
</ol> 4。体外循环与血气分析 <ol> 用于血气分析的 <li> , 使用玻璃毛细管收集 60-90 和 #181; 通过股动脉导管进行动脉血流. 
	<ol> <li> 在硅胶导管上使用一个小钳, 并将导管与导尿管分离。在钳上缓慢释放压力, 使毛细管控制充填. </li>
		<li> 将硅胶管重新连接到导管。将毛细管插入血气分析仪, 用于测量在以下时间点: 10 分钟后启动 CPB 通气 (BGA1)25分钟的 cpb 和15分钟的呼吸骤停 (BGA2) 后40分钟 cpb 和30分钟的呼吸骤停 (BGA3) 后55分钟, 45 分钟的呼吸, 20 分钟心跳骤停 (BGA4) </li> </ol> </li> <li> 在稳定的 CPB 流下, initia通过停止通气来呼吸骤停. </li>
	<li> 通风结束后, 将暖垫设置为28和 #186; c 和 #160; 开始局部冷却的动物到28和 #186; c 体温在第一20分钟内使用纱布浸泡在生理盐水中的呼吸骤停. </li>
	<li> 一旦体温为28和 #186; C 是达到, 并在总共30分钟的呼吸骤停后, 管理0.3 毫升的7.45% 氯化钾溶液进入 CPB 回路, 启动心脏停搏液. </li>
	<li> 对于主动脉的交叉夹紧, 将先前放置的丝环 (步骤 3.10) 置于颅骨方向, 以便更好地曝光, 并在主动脉的上升部分放置微 serrefine 钳. </li>
	<li> 共执行60分钟呼吸骤停和30分钟心跳骤停。从提升主动脉中取出微 serrefine 钳, 开始心脏再灌注。同时, re-ventilate 和温暖的动物到37和 #176; C. </li>
	<li> 在再灌注完成后, 动物已达到常温, 关闭泵, 并继续监测生理测量 ( 如 体温、心电图和侵入性血压) 20 分钟, 终止 CPB。
	<li> 在实验结束时, 血完全麻醉下的动物 (5% 异氟醚), 并收集血液和器官作进一步分析 14 . </li>
</ol>

<ol>
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作者没有什么可透露的。

我们已经开发了一个完全功能性的临床相关模型的 CPB 在鼠标。由于有三十多株患有心血管疾病的小鼠, 我们的模型可以成为与 CPB 相关的新的前瞻性协议的发展起点。此外, 由于大量的鼠标特异试剂和 knockout-out 小鼠, 该模型不仅可以取代目前的大鼠 cpb 模型, 但将有助于解剖的分子机制参与与 cpb 相关的器官损害。到目前为止, CPB 还没有应用于小鼠, 因为在插管技术的显微外科挑战, 以及技术挑战, ?...

自体外循环 (cpb) 进入临床后, 它在心脏手术中起了重要作用1。在现代心脏手术中, 延长的 CPB 时间对进行广泛的主动脉重建和联合手术是必不可少的。尽管技术进步巨大, 外循环的使用与术后全身和局部脏器损伤有关2,3。
开发了大型动物模型来研究 CPB 在生理过程中的作用4,5。虽然这些模型提供了一些对 CPB 相关并发症的洞察力, 但它们是非常昂贵的, 分子工具 (例如, 抗体) 是非常有限的。一个更具成本效益的替代品已经在小动物身上得到了发展。自其发展以来, 进行了多项研究, 以优化大鼠和家兔的 CPB 模型5,6,7,8,9。这些模型为病理生理学疾病过程的测量提供了良好的基础;然而, 由于缺乏相关的抗体和试剂, 它们仍然不足以研究细胞和体液免疫学。这削弱了他们在这个研究领域的作用。
我们最近开发了一个鼠 CPB 模型。由于各种鼠标特异试剂和转基因小鼠, 小鼠模型一般是生理、分子和免疫学研究的选择模型10,11。因此, 我们的模型将促进研究 CPB 与各种并存, 因为有许多小鼠菌株可与临床相关的疾病12,13。因此, 本文详细介绍了如何在小鼠体内进行 CPB。在深部呼吸和循环停止后, 对氧和血流动力学参数进行严密监测。

<table><tbody><tr><td>Sterofundin</td><td>B.Braun Petzold GmbH</td><td>PZN:8609189</td><td>priming volume, 1:1 with Tetraspan</td></tr><tr><td>Tetraspan 6% HES Solution</td><td>B. Braun Melsungen AG</td><td>PZN: 05565416</td><td>priming volume, 1:1 with Sterofundin</td></tr><tr><td>Heparin Natrium 25.000</td><td>Ratiopharm GmbH</td><td>PZN: 3029843</td><td>2.5 IU per ml of priming solution</td></tr><tr><td>NaHCO&lt;sub&gt;3&lt;/sub&gt; 8,4% Solution</td><td>B. Braun Melsungen AG</td><td>PZN: 1579775</td><td>3% in priming solution</td></tr><tr><td>KCL 7.45 % Solution</td><td>B. Braun Melsungen AG</td><td>PZN: 2418577</td><td>0.1 mL for cardioplegia</td></tr><tr><td>Carprofen</td><td>Zoetis Inc., USA</td><td>PZN:00289615 08859153</td><td>5 mg/kg/BW</td></tr><tr><td>1 Fr PU Catheter</td><td>Instechlabs INC., USA</td><td>C10PU-MCA1301</td><td>carotid artery</td></tr><tr><td>2 Fr PU Catheter</td><td>Instechlabs INC., USA</td><td>C20PU-MJV1302</td><td>jugular vein</td></tr><tr><td>Vasofix Safety catheter 20G</td><td>B.Braun Medical</td><td>4268113S-01</td><td>orotracheal intubation</td></tr><tr><td>8-0 Silk suture braided</td><td>Ashaway Line &amp;amp; Twine Mfg. Co., USA</td><td>75290</td><td>ligature</td></tr><tr><td>Isoflurane</td><td>Piramal Critical Care Deutschland GmbH</td><td>PZN:9714675</td><td>narcosis</td></tr><tr><td>CLINITUBES blood capillaries</td><td>Radiomed GmbH</td><td>51750132</td><td>blood sampling 60 - 95 microliter</td></tr><tr><td>Spring Scissors - 6mm Blades</td><td>Fine Science Tools GmbH</td><td>15020-15</td><td>instruments</td></tr><tr><td>Spring Scissors - 2mm Blades</td><td>Fine Science Tools GmbH</td><td>15000-03</td><td>instruments</td></tr><tr><td>Halsted-Mosquito Hemostat</td><td>Fine Science Tools GmbH</td><td>13009-12</td><td>instruments</td></tr><tr><td>Dumont #55 Forceps</td><td>Fine Science Tools GmbH</td><td>11295-51</td><td>instruments</td></tr><tr><td>Castroviejo Micro Needle Holder - 9cm</td><td>Fine Science Tools GmbH</td><td>12060-02</td><td>instruments</td></tr><tr><td>Micro Serrefines</td><td>Fine Science Tools GmbH</td><td>18555-01</td><td>instruments</td></tr><tr><td>Bulldog Serrefine</td><td>Fine Science Tools GmbH</td><td>18050-28</td><td>instruments</td></tr><tr><td>MiniVent Ventilator for Mice (Model 845)</td><td>Harvard Apparatus</td><td>73-0044</td><td>mechanical ventilation</td></tr><tr><td>Isoflurane Vaporizer Drager 19.1</td><td>Dr&amp;auml;gerwerk AG &amp;amp; Co. KGaA</td><td></td><td>anesthesia 1.3 - 2.5%</td></tr><tr><td>PowerLab data acquisition device 4/35</td><td>ADInstruments Ltd, New Zealand</td><td>PL3504</td><td>invasive pressure, ECG, temperature</td></tr><tr><td>ABL 800 Flex</td><td>Radiometer GmbH</td><td></td><td>blood gas analysis</td></tr><tr><td>NMRI mice</td><td>Charles River Laboratories</td><td>Crl:NMRI(Han)</td><td>male, 30 - 35 g, 12 weeks old, housed at least 1 week before the experiment</td></tr></tbody></table>

cardiopulmonary bypass in a mouse model: a novel approach

随着长期体外循环在心脏介入过程中变得越来越重要, 临床需求增加, 以进行程序优化, 并最大限度地减少因长期外循环而造成的器官损害。本文的目的是演示一个完整的功能和临床相关模型的体外循环在小鼠。我们报告的设备设计, 灌注电路优化, 和显微外科技术。该模型是一个急性模型, 这是不相容的生存, 由于需要多血图。由于可用于小鼠的工具范围 (例如、标记、敲除、等), 该模型将有助于研究器官损伤的分子机制和体外循环对其他并存.

本协议描述了在鼠 CPB 过程中的灌注回路、手术过程和生理参数的监测。当由一个充分熟练的 microsurgeon, 结果是一致和性获得。
为了保持足够的组织灌注, 平均动脉压力始终保持在40和60柱通过调整 CPB 血流量和增加额外的体积。根据动物的重量, 它的体积状况和体温, 外血流维持在 2.3-6.5 毫升/分钟. 在实验过程中, 通过增加0.1 毫...

Watch this Scientific Journal Video about Cardiopulmonary Bypass in a Mouse Model: A Novel Approach at JoVE.com

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

本文介绍了如何在小鼠体内进行体外循环手术。这种新型模型将有助于研究器官损伤的分子机制。

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 ...

cardiopulmonary-bypass-in-a-mouse-model-a-novel-approach

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

10.8K 次观看.  Hannover Medical School. 本文介绍了如何在小鼠体内进行体外循环手术。这种新型模型将有助于研究器官损伤的分子机制。

视频: Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

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 ...

科研

医学

A Modified Method for Heterotopic Mouse Heart Transplantion

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 available. A difficulty is presented due to the small size of the animal and the considerable technical challenges of the microsurgery involved in heart transplantation. In particular, a high rate of technical failure early after transplantation may result from recipient death and post-operative complications such as hind limb paralysis or a non-beating heart. Here, the complete technique for heterotopic mouse heart transplantation is demonstrated, involving harvesting the donor heart and its subsequent implantation into a recipient mouse. The donor heart is harvested immediately following in situ perfusion with cold heparinized saline and transection of the ascending aorta and pulmonary artery. The recipient operation involves preparation of the abdominal aorta and inferior vena cava (IVC), followed by end-to-side anastomosis of the donor aorta with the recipient aorta using a single running 10-0 microsuture and a similar anastomosis of the donor pulmonary artery with the recipient IVC. Following the operation the animal is injected with 0.6 ml normal saline subcutaneously and allowed to recover on a 37&#160;&#176;C heating pad. The results from 227 mouse heart transplants are summarized with a success rate at 48 hr of 86.8%. Of the 13.2% failures within 48 hr, 5 (2.2%) experienced hind limb paralysis, 10 (4.4%) had a non-beating heart due to graft ischemic injury and/or thrombosis, while 15 (6.6%) died within 48 hr.

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 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 Recovery Cardiopulmonary Bypass Model Without Transfusion or Inotropic Agents in Rats

Cardiopulmonary bypass (CPB) is indispensable in cardiovascular surgery. Despite the dramatic refinement of CPB technique and devices, multi-organ complications related to prolonged CPB still compromise the outcome of cardiovascular surgeries, and may worsen postoperative morbidity and mortality. Animal models recapitulating the clinical usage of CPB enable the clarification of the pathophysiological processes that occur during CPB, and facilitate pre-clinical studies to develop strategies protecting against these complications. Rat CPB models are advantageous because of their greater cost-effectiveness, convenient experimental processes, abundant testing methods at the genetic or protein levels, and genetic consistency. They can be used for investigating the immune system activation and synthesis of proinflammatory cytokines, compliment activation, and production of oxygen free radicals. The rat models have been refined and have gradually taken the place of large-animal models. Here, we describe a simple CPB model without transfusion and/or inotropic agents in a rat. This recovery model allows the study of the long-term multiple organ sequelae of CPB.

Here, we present a protocol to describe a simple recovery cardiopulmonary bypass model without transfusion or inotropic agents in a rat. This model allows the study of the long-term multiple organ sequelae of cardiopulmonary bypass.

无输血或强心剂剂的大鼠心肺旁路移植模型

Cardiopulmonary bypass (CPB) is indispensable in cardiovascular surgery. Despite the dramatic refinement of CPB technique and devices, multi-organ ...

免疫与感染

Transplantation of a 3D Bioprinted Patch in a Murine Model of Myocardial Infarction

Testing regenerative properties of 3D bioprinted cardiac patches in vivo using murine models of heart failure via permanent left anterior descending (LAD) ligation is a challenging procedure and has a high mortality rate due to its nature. We developed a method to consistently transplant bioprinted patches of cells and hydrogels onto the epicardium of an infarcted mouse heart to test their regenerative properties in a robust and feasible way. First, a deeply anesthetized mouse is carefully intubated and ventilated. Following left lateral thoracotomy (surgical opening of the chest), the exposed LAD is permanently ligated and the bioprinted patch transplanted onto the epicardium. The mouse quickly recovers from the procedure after chest closure. The advantages of this robust and quick approach include a predicted 28-day mortality rate of up to 30% (lower than the 44% reported by other studies using a similar model of permanent LAD ligation in mice). Moreover, the approach described in this protocol is versatile and could be adapted to test bioprinted patches using different cell types or hydrogels where high numbers of animals are needed to optimally power studies. Overall, we present this as an advantageous approach which may change preclinical testing in future studies for the field of cardiac regeneration and tissue engineering.

This protocol aims to transplant a 3D bioprinted patch onto the epicardium of infarcted mice modeling heart failure. It includes details regarding anesthesia, the surgical chest opening, permanent ligation of the left anterior descending (LAD) coronary artery and application of a bioprinted patch onto the infarcted area of the heart.

心肌梗塞穆林模型中的 3D 生物打印贴片移植

Testing regenerative properties of 3D bioprinted cardiac patches in vivo using murine models of heart failure via permanent left anterior descending (LAD) ...

生物工程

Optimization of the Cuff Technique for Murine Heart Transplantation

Murine cardiac transplantation has been performed for more than 40 years. With advancements in microsurgery, certain new techniques have been used to improve surgical efficiency. In our lab, we have optimized the cuff technique with two major steps. First, we used the inner tube technique to insert a temporary inner tube into the external jugular vein and carotid artery blood vessel to facilitate eversion of the vessel over the cuff. Second, we performed complete heterotopic cardiac transplantation through the collaboration of two experienced surgeons. These modifications effectively reduced the operation time to 25 minutes, with a success rate of 95%. In this report, we describe these procedures in detail and provide a supplemental video. We believe that this report on the improved cuff technique will offer practical guidance for murine heterotopic heart transplantation and will enhance the utility of this mouse model for basic research.

We introduce an inner tube approach to the cuff technique for mouse cervical heterotopic heart transplantation to help evert the vessel over the cuff. We found that cooperation between two experienced surgeons markedly shortens the operation time.

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 ...

Veno-Venous Extracorporeal Membrane Oxygenation in a Mouse

The use of extracorporeal membrane oxygenation (ECMO) has increased substantially in recent years. ECMO has become a reliable and effective therapy for acute as well as end-stage lung diseases. With the increase in clinical demand and prolonged use of ECMO, procedural optimization and prevention of multi-organ damage are of critical importance. The aim of this protocol is to present a detailed technique of veno-venous ECMO in a non-intubated, spontaneously breathing mouse. This protocol demonstrates the technical design of the ECMO and surgical steps. This murine ECMO model will facilitate the study of pathophysiology related to ECMO (e.g., inflammation,bleeding and thromboembolic events). Due to the abundance of genetically modified mice, the molecular mechanisms involved in ECMO-related complications can also be dissected.

Here we present a protocol describing the technique of veno-venous extracorporeal membrane oxygenation (ECMO) in a non-intubated, spontaneously breathing mouse. This murine model of ECMO can be effectively implemented in experimental studies of acute and end-stage lung diseases.

小鼠静脉内膜氧化术

The use of extracorporeal membrane oxygenation (ECMO) has increased substantially in recent years. ECMO has become a reliable and effective therapy for ...

Mouse Heterotopic Cervical Cardiac Transplantation Utilizing Vascular Cuffs

Murine models of cardiac transplantation are frequently utilized to study ischemia-reperfusion injury, innate and adaptive immune responses after transplantation, and the impact of immunomodulatory therapies on graft rejection. Heterotopic cervical heart transplantation in mice was first described in 1991 using sutured anastomoses and subsequently modified to include cuff techniques. This modification allowed for improved success rates, and since then, there have been multiple reports that have proposed further technical improvements. However, translation into more widespread utilization remains limited due to the technical difficulty associated with graft anastomoses, which requires precision to achieve adequate length and caliber of the cuffs to avoid vascular anastomotic twisting or excessive tension, which can result in damage to the graft. The present protocol describes a modified technique for performing heterotopic cervical cardiac transplantation in mice which involves cuff placement on the recipient's common carotid artery and the donor's pulmonary artery in alignment with the direction of the blood flow.

Mouse cardiac transplantation models represent valuable research tools for studying transplantation immunology. The present protocol details mouse heterotopic cervical cardiac transplantation that involves the placement of cuffs on the recipient's common carotid artery and the donor's pulmonary artery trunk to allow for laminar blood flow.

利用血管袖带的小鼠异位宫颈心脏移植

Murine models of cardiac transplantation are frequently utilized to study ischemia-reperfusion injury, innate and adaptive immune responses after ...

Blood Circuit Reconstruction in an Abdominal Mouse Heart Transplantation Model

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the established technique for a modified blood circuit reconstruction in a heterotopic abdominal heart transplantation model is presented. This method uses the intrathoracic inferior vena cava (IIVC) instead of the pulmonary artery of the donor heart for the anastomosis to the inferior vena cava of the recipient. It is facilitating and improving success rates for abdominal heart transplantation in mice.

A novel technique for blood circuit reconstruction in a heterotopic abdominal mouse heart transplantation model is demonstrated.

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 ...

A Modified Cuff Technique for Mouse Cervical Heterotopic Heart Transplantation Model

Cardiac allograft rejection limits the long-term survival of patients after heart transplantation. A mouse heart transplantation model is ideal for investigating the mechanism of cardiac allograft rejection in preclinical studies because of their high homology with human genes. This understanding would help develop unique approaches to improving patients&#39; long-term survival treated with cardiac allografts. In a mouse model, abdominal donor heart implantation is commonly performed with an end-to-end&#160;anastomosis to the recipient&#39;s aorta and inferior vena cava using stitches. In this model, the donor&#39;s heart is implanted by end-to-end anastomosis to the recipient&#39;s carotid artery and jugular vein by the modified-Cuff technique. The transplantation surgery is performed without stitching and thus may increase the survival of the recipient since there is no interference with the blood supply and venous reflux of the lower body. This mouse model would help investigate the mechanisms underlying the immunological and pathological (acute/chronic) rejection of cardiac allografts.

In the present protocol, a mouse heart transplantation model is used for investigating the mechanism of cardiac allograft rejection. In this heterotopic heart transplantation model, operation efficiency is improved, and the survival of cardiac grafts is ensured by a cervical end-to-end anastomosis of heart implantation using a modified Cuff technique.

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 ...