The authors would like to thank the many individuals who have worked on and perfected this technique and have been responsible for the generation of many manuscripts both in this lab and others. This work was supported by National Institutes of Health Grant EY12707 (PMS) and an unrestricted grant from Research to Prevent Blindness to Department of Ophthalmology.

注意：在此过程中使用的所有动物都按照研究协会在视觉与眼科语句在眼科和视觉研究用动物，以及订定圣路易斯大学动物监督委员会的指导方针进行处理。 注：所有手术器械和解决方案之前进行消毒手术，以限制眼睛微生物感染。应当指出的是，虽然动物做这个过程遇到一些痛苦，我们不使用止痛剂。这样做的理由是因为所有的止痛药是消炎和自角膜移植反应涉及炎症，使用抗炎药物会损害我们确定参与角膜移植失败哪些因素的能力。 1.麻醉<ol style="margin-left: 40px;"><li>氯胺酮（86腹腔注射将在全身麻醉供体和受体的小鼠。98毫克/公斤）和甲苯噻嗪（13.04毫克/千克）。 </li><li>保持麻醉状态下的受体小鼠在整个这通常需要30分钟到一个小时的过程。因此，不断地监视鼠标苏醒的迹象。 </li><li>申请puralube软膏，不会接受手术后的动物被麻醉，以防止干眼。 </li></ol> 2.角膜移植术<ol style="margin-left: 40px;"><li>获得捐赠的角膜按钮。 <ol><li>一旦动物完全麻醉，达到充分散瞳由一对夫妇1％托吡卡胺和2.5％盐酸去氧肾上腺素的眼药水管理。 </li><li>放置供体动物的头部水平在电路板上放置在一个坚固的可移动的支持。固定头的整个颈部带的条带，以保证眼睛处于整个操作的水平位置。 </li><li>使用2毫米直径的环钻，其前端被染成用甲基蓝，出行角膜中央部位移植。 </li><li>用锋利的刀片，穿透角膜与注入healon进入前房加深它减少损坏施主内皮和底层透镜的机会。 </li><li>切除供体移植物与vannas剪刀，并放入含有Hanks平衡盐溶液，直到使用一盘菜。 </li><li>后供体移植物已被移除，通过安乐死二氧化碳吸入供体小鼠。 </li></ol></li><li>准备移植床。 <ol><li>如在2.1.1中描述过2.1.2收件人重复相同的步骤。 </li><li>使用1.5毫米直径的环钻，勾勒出收件人移植部位。 </li><li>用锋利的刀片，穿透角膜与注入healon进入前房加深它来减少损伤的机会底层透镜。 </li><li>使用vannas剪刀取下收件人概述角膜中央按钮并丢弃。 </li></ol></li><li>缝合接枝<ol><li>将供体角膜在移植床收件人的角膜。确保有足够的healon是在角膜供体，以保护捐赠者的内皮细胞通过镜头直接接触伤害。 </li><li>使用超细尖microforceps，放置11-0尼龙缝线咬头到捐助方，通过与全层到收件人的面为90％深度的捐助，然后打结。 </li><li>一旦角膜被固定到位，执行midcardinal间断缝合，使得所述角膜有8至10个总缝线和供体角膜牢固地排队与和连接到接收方的角膜移植物床。 </li></ol></li><li>深化前房<ol><li>通过注入的HBSS或气泡进入前房加深前房并轻轻检查角膜移植物的完整性对于泄漏用纤维素海绵。 注意：如果前房不能改革则是C的概率高ataract这将使得移植角膜的未来的评估很困难，也将有可能导致供体角膜内皮功能紊乱，因此移植物衰竭。 </li></ol></li><li>终期评估<ol><li>观察的眼睛，以确定瞳孔是圆的，前房深度是正常的。 注意：如果瞳孔不圆这表明缝合虹膜过程中损坏，因此，移植被认为是一个技术故障。 </li><li>应用抗生素软膏的眼睛。可选：合上眼睑用7-0丝线缝合。 </li><li>观察小鼠直到它们完全清醒，然后分别容纳他们最少两天后手术。 </li></ol></li></ol> 3.拆线<ol style="margin-left: 40px;"><li>在盖体缝合用于这些情况下，麻醉上述老鼠和盖子取下缝合处48小时。 </li><li>麻醉小鼠术后7天。取出suturES确保角膜移植。一旦缝线被删除，动物已经完全觉醒，将其放回笼子。 </li></ol> 4.临床评价<ol style="margin-left: 40px;"><li>检查眼适应症手术并发症，包括，白内障（晶状体混浊），前房出血（血液中的前房）的前房，它是适当的深度，或角膜的显著不透明度不。想想那些表现出这些复杂的“妥协”，并通过二氧化碳吸入安乐死他们。 <ol><li>执行对未麻醉的小鼠的所有考试。按住鼠标，用一只手从而抑制鼠标，这样另一方面可以proptose眼睛，使眼睛更好的视野。一旦观测完成后，返回到动物的笼子。 </li></ol></li><li>有一个观察者不熟悉治疗组评估所移植的角膜每周2至3次，角膜移植的迹象排斥或角膜移植失败。请使用手术显微镜或水平裂隙灯生物显微镜对这些意见。 <ol><li>评估用于使用比例为0〜5的不透明度的尺度定义如下每角膜： <ol><li>分配给0分给那些有混浊的迹象眼角膜。 </li><li>指定1分给那些表现出最小的浅表角膜混浊。 </li><li>分配给2分，以用于显示轻度和更深的不透明性，但潜在的瞳孔和虹膜仍是有迹可寻的眼角膜。 </li><li>分配3分至角膜中显示基质的不透明度，其特征在于所述虹膜不能看到详细除瞳孔边缘的。 </li><li>分配的4分至角膜中显示致密的基质混浊并且如果没有下面的结构能够被观看。 </li><li>分配给5分，以角膜中显示完整的不透明性和密集的间质水肿，瞳孔与虹膜完全遮蔽。 </li></ol></li><li><html>还评估每个角膜供使用的1〜8的等级规模要做到这一点的血管浸润（新血管形成）的程度，查看角膜作为组成4相等象限并确定在每一个象限中具有的血管的量分数，范围从0（无船），以2象限广泛的血管。添加单独得分从各象限来计算最终新生血管得分。 </li><li>归类为急性排斥，如果他们有连续两次观测的比分为3点的时间最多5周的眼角膜。 </li><li>分类小鼠的角膜是透明的5个星期，但发展混浊，有时&gt;45天植入后，用连续两个时间点分3，具有经过后期长期的角膜移植排斥反应。采用Kaplan-Meier生存曲线分析移植物的存活。 </li></ol></li></ol> 5.操纵的模型在左：40PX;“&gt; <li>制备单细胞脾。 <ol><li>制备单细胞，首先安乐死的供体小鼠。然后，取出脾脏。 </li><li>将脾脏在细胞过滤并破坏其与由3毫升注射器的注射器柱塞。 </li><li>在10毫升Hanks平衡盐溶液中洗涤细胞并重新悬浮。 </li><li>删除10微升细胞悬液，加入10微升0.4％台盼蓝混匀。补充一点，对血球和细胞计数在中央格。细胞在管的数量是细胞数目×10 4×2（稀释因子中的台盼蓝）×10（在管体积）。 </li></ol></li><li>注射进入前房。 <ol><li>麻醉小鼠，如前所述。 </li><li>执行使用解剖显微镜注射。对于每个房内注射，使用10 6脾细胞中0.005毫升容积并配有一个33克针一0.25毫升微升注射器。 注意：其他操作该模型可通过治疗动物与充当要么拮抗剂或激动剂，以确定特定因素可能起以下原位角膜移植术中的作用的试剂进行。 </li></ol></li></ol>

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K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Presurgical+corticosteroid+treatment+improves+corneal+transplant+survival+in+mice.">Presurgical corticosteroid treatment improves corneal transplant survival in mice.</a> Cornea. 32, 1591-1598 (2013).</li><li>Yamazoe, K., Yamazoe, K., Shimazaki-Den, S., Shimazaki, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prognostic+factors+for+corneal+graft+recovery+after+severe+corneal+graft+rejection+following+penetrating+keratoplasty.">Prognostic factors for corneal graft recovery after severe corneal graft rejection following penetrating keratoplasty.</a> BMC Ophthalmol. 13, 5 (2013).</li><li>Panda, A., Vanathi, M., Kumar, A., Dash, Y., Priya, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Corneal+graft+rejection.">Corneal graft rejection.</a> Surv. Ophthalmol. 52, 375-396 (2007).</li><li>Patel, S. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Graft+survival+and+endothelial+outcomes+in+the+new+era+of+endothelial+keratoplasty.">Graft survival and endothelial outcomes in the new era of endothelial keratoplasty.</a> Exp. Eye Res. 95, 40-47 (2012).</li><li>Anshu, A., Price, M. O., Tan, D. T., Price, F. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+keratoplasty:+a+revolution+in+evolution.">Endothelial keratoplasty: a revolution in evolution.</a> Surv. Ophthalmol. 57, 236-252 (2013).</li></ol>

The authors have no competing financial interests.

这里描述角膜移植的小鼠模型使研究人员研究人类角膜移植排斥反应的模型，该模型预测的因素是什么，最好既排斥15,17,18,20，26-30和角膜的验收21-25关联移植。不象人类角膜移植，其中给予患者或者局部或全身激素治疗，以任一治疗或预防排斥31，该模型通常用于确定那些是相关的同种异体移植排斥反应在不存在这样的治疗的因素。除了一款型号为急性角膜移植排斥反应?...

角膜移植是最成功的和常见的类型移植在人执行的一个。为什么进行该手术的原因是损伤，感染性疾病1，或其他形式的非感染性角膜病2的结果。从美国眼库协会的数据显示，46000在2011年进行（见网站：restoresight.org/eye_banks/eye_banks.html）。其成功的标志是有一年不合格率同种异体角膜移植物的范围从10％至15％，并在5年的成功是超过70％3-8。因为许多研究已经显示，角膜移植的成功是直接关系到一个事实，即眼睛的免疫特权部位。负责角膜状态作为免疫特权部位因素包括缺乏既血管和淋巴管在角膜中，相对缺乏抗原呈递细胞的因子，由角膜该suppres产生免疫效应器funtions 9-15，MHC的低表达抗原16，和FasL 17-20的表达。 然而，尽管这些因素诱发这些移植成功的，它们经历排斥3-7。因此，了解这些调解这种排斥反应的机制，以及测试各种治疗，以防止排斥反应是至关重要的。为此，我们在这里描述角膜移植是已经使用了超过20年，研究角膜移植在受控的实验环境的小鼠模型。自移植反应涉及协同工作的许多不同的因素，这将最终确定移植的组织是否失败或成功，则它不可能了解这些因素中的任何体外模型中的重要性。因此，使用完整的动物研究是必需的，以确定哪些因素是重要的成功或者failu重新移植组织。 而其它种类的动物被用来研究角膜移植中，当相比于使用其他种类的鼠模型中具有几个优点。第一个是表示某些转基因的或已被基因靶向至缺乏特异性免疫因子，其功能在移植可以更好地研究了表达小鼠的许多菌株的存在。此外，还有许多试剂（包括重组因子和抗体是中和因子）是特定于小鼠，并且不为其他许多种动物的存在。由于这些因素的存在，该模型已被广泛用于鉴定参与急性角膜移植反应15，17,18,20 -29有关因素。此外，有许多参与角膜移植的因素也是已知的是功能在其他组织中的移植。

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			<td height="21" style="height:21px;width:216px;">Zeiss Surgical Microscope</td>
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</table>

murine corneal transplantation: a model to study the most common form of solid organ transplantation

Corneal transplantation is the most common form of organ transplantation in the United States with between 45,000 and 55,000 procedures performed each year. While several animal models exist for this procedure and mice are the species that is most commonly used. The reasons for using mice are the relative cost of using this species, the existence of many genetically defined strains that allow for the study of immune responses, and the existence of an extensive array of reagents that can be used to further define responses in this species. This model has been used to define factors in the cornea that are responsible for the relative immune privilege status of this tissue that enables corneal allografts to survive acute rejection in the absence of immunosuppressive therapy. It has also been used to define those factors that are most important in rejection of such allografts. Consequently, much of what we know concerning mechanisms of both corneal allograft acceptance and rejection are due to studies using a murine model of corneal transplantation. In addition to describing a model for acute corneal allograft rejection, we also present for the first time a model of late-term corneal allograft rejection.

已用于角膜移植的小鼠模型中超过20年，成功刻画两个角膜移植排斥反应19-23和角膜移植验收13，15,16,18，24-27机制。该模型被用于建立在角膜移植接受FasL表达的重要性，在该动物缺乏FasL的不能够接受角膜移植物15。它也被用来表明，血管内皮生长因子受体1啉治疗显著增加角膜移植存活28。在一个非常最近的一份报告此模型来之前，角膜移植测试是否小鼠的预处理与...

Watch this Scientific Journal Video about Murine Corneal Transplantation: A Model to Study the Most Common Form of Solid Organ Transplantation at JoVE.com

Murine Corneal Transplantation: A Model to Study the Most Common Form of Solid Organ Transplantation

Mice have been used as a model for studying many forms of transplantation, including corneal transplantation. We describe in this report a murine model for both acute and late-term corneal transplantation.

小鼠角膜移植模型，研究实体器官移植的最常见形式

Corneal transplantation is the most common form of organ transplantation in the United States with between 45,000 and 55,000 procedures performed each ...

murine-corneal-transplantation-model-to-study-most-common-form-solid

Research

JoVE Journal

Medicine

9.6K 次观看.  Saint Louis University. Mice have been used as a model for studying many forms of transplantation, including corneal transplantation. We describe in this report a murine model for both acute and late-term corneal transplantation.

视频: Murine Corneal Transplantation: A Model to Study the Most Common Form of Solid Organ Transplantation

Orthotopic Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant vasculopathy (TVP).
The commonly used animal model for cardiovascular chronic rejection studies is the heterotopic heart transplant model performed in laboratory rodents. This model is used widely in experiments since Ono and Lindsey (3) published their technique. To analyze the findings in the blood vessels, the heart has to be sectioned and all vessels have to be measured.
Another method to investigate chronic rejection in cardiovascular questionings is the aortic transplant model (1, 2). In the orthotopic aortic transplant model, the aorta can easily be histologically evaluated (2). The PVG-to-ACI model is especially useful for CAV studies, since acute vascular rejection is not a major confounding factor and Cyclosporin A (CsA) treatment does not prevent the development of CAV, similar to what we find in the clinical setting (4). A7-day period of CsA is required in this model to prevent acute rejection and to achieve long-term survival with the development of TVP.
This model can also be used to investigate acute cellular rejection and media necrosis in xenogeneic models (5).

This video demonstrates the orthotopic aortic transplant model as a simple model to study the development of transplant vasculopathy (TVP) in rats.

主动脉原位移植大鼠模型，研究慢性血管病变的发展

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant ...

科研

医学

A Method for Murine Islet Isolation and Subcapsular Kidney Transplantation

Since the early pioneering work of Ballinger and Reckard demonstrating that transplantation of islets of Langerhans into diabetic rodents could normalize their blood glucose levels, islet transplantation has been proposed to be a potential treatment for type 1 diabetes 1,2. More recently, advances in human islet transplantation have further strengthened this view 1,3. However, two major limitations prevent islet transplantation from being a widespread clinical reality: (a) the requirement for large numbers of islets per patient, which severely reduces the number of potential recipients, and (b) the need for heavy immunosuppression, which significantly affects the pediatric population of patients due to their vulnerability to long-term immunosuppression. Strategies that can overcome these limitations have the potential to enhance the therapeutic utility of islet transplantation.
Islet transplantation under the mouse kidney capsule is a widely accepted model to investigate various strategies to improve islet transplantation. This experiment requires the isolation of high quality islets and implantation of islets to the diabetic recipients. Both procedures require surgical steps that can be better demonstrated by video than by text. Here, we document the detailed steps for these procedures by both video and written protocol. We also briefly discuss different transplantation models: syngeneic, allogeneic, syngeneic autoimmune, and allogeneic autoimmune.

Transplantation of isolated islets has been proposed to be a potential treatment for type 1 diabetes. Here we describe a method to isolate islets from mouse pancreata and transplant them to the subcapsular space of the kidney.

小鼠胰岛的分离和包膜下肾移植的方法

Since the early pioneering work of Ballinger and Reckard demonstrating that transplantation of islets of Langerhans into diabetic rodents could normalize ...

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 ...

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 ...

Cardiac Catheterization in Mice to Measure the Pressure Volume Relationship: Investigating the Bowditch Effect

Animal models that mimic human cardiac disorders have been created to test potential therapeutic strategies. A key component to evaluating these strategies is to examine their effects on heart function. There are several techniques to measure in vivo cardiac mechanics (e.g., echocardiography, pressure/volume relations, etc.). Compared to echocardiography, real-time left ventricular (LV) pressure/volume analysis via catheterization is more precise and insightful in assessing LV function. Additionally, LV pressure/volume analysis provides the ability to instantaneously record changes during manipulations of contractility (e.g., &#946;-adrenergic stimulation) and pathological insults (e.g., ischemia/reperfusion injury). In addition to the maximum (+dP/dt) and minimum (-dP/dt) rate of pressure change in the LV, an accurate assessment of LV function via several load-independent indexes (e.g., end systolic pressure volume relationship and preload recruitable stroke work) can be attained. Heart rate has a significant effect on LV contractility such that an increase in the heart rate is the primary mechanism to increase cardiac output (i.e., Bowditch effect). Thus, when comparing hemodynamics between experimental groups, it is necessary to have similar heart rates. Furthermore, a hallmark of many cardiomyopathy models is a decrease in contractile reserve (i.e., decreased Bowditch effect). Consequently, vital information can be obtained by determining the effects of increasing heart rate on contractility. Our and others data has demonstrated that the neuronal nitric oxide synthase (NOS1) knockout mouse has decreased contractility. Here we describe the procedure of measuring LV pressure/volume with increasing heart rates using the NOS1 knockout mouse model.

This article describes the measurement of murine left ventricular function via pressure/volume analysis at different heart rates.

心导​​管在小鼠测量压力体积的关系：调查的影响鲍迪奇

Animal models that mimic human cardiac disorders have been created to test potential therapeutic strategies. A key component to evaluating these ...

A Porcine Corneal Endothelial Organ Culture Model Using Split Corneal Buttons

Experimental research on corneal endothelial cells is associated with several difficulties. Human donor corneas are scarce and rarely available for experimental investigations as they are normally needed for transplantation. Endothelial cell cultures often do not translate well to in vivo situations. Due to the biostructural characteristics of non-human corneas, stromal swelling during cultivation induces substantial corneal endothelial cell loss, which makes it difficult to perform cultivation for an extended period of time. Deswelling agents such as dextran are used to counteract this response. However, they also cause significant endothelial cell loss. Therefore, an ex vivo organ culture model not requiring deswelling agents was established. Pig eyes from a local slaughterhouse were used to prepare split corneal buttons. After partial corneal trephination, the outer layers of the cornea (epithelium, bowman layer, parts of the stroma) were removed. This significantly reduces corneal endothelial cell loss induced by massive stromal swelling and Descemet&#39;s membrane folding throughout longer cultivation periods and improves general preservation of the endothelial cell layer. Subsequent complete corneal trephination was followed by the removal of the split corneal button from the remaining eye bulb and cultivation. Endothelial cell density was assessed at follow-up times of up to 15 days after preparation (i.e., days 1, 8, 15) using light microscopy. The preparation technique used allows a better preservation of the endothelial cell layer enabled by less stromal tissue swelling, which results in slow and linear decline rates in split corneal buttons comparable to human donor corneas. As this standardized organo-typically cultivated research model for the first time allows a stable cultivation for at least two weeks, it is a valuable alternative to human donor corneas for future investigations of various external factors with regards to their effects on the corneal endothelium.

Here, a step-by-step protocol for the preparation and cultivation of porcine split corneal buttons is presented. As this organo-typically cultivated organ culture model shows cell death rates within 15 days, comparable to human donor corneas, it represents the first model allowing long-term cultivation of non-human corneas without adding toxic dextran.

使用分裂的角质按钮的孔膜内皮器官培养模型

Experimental research on corneal endothelial cells is associated with several difficulties. Human donor corneas are scarce and rarely available for ...

Development of a Noninvasive, Laser-Assisted Experimental Model of Corneal Endothelial Cell Loss

Nd:YAG lasers have been used to perform noninvasive intraocular surgery, such as capsulotomy for several decades now. The incisive effect relies on the optical breakdown at the laser focus. Acoustic shock waves and cavitation bubbles are generated, causing tissue rupture. Bubble sizes and pressure amplitudes vary with pulse energy and position of the focal point. In this study, enucleated porcine eyes were positioned in front of a commercially available Nd:YAG laser. Variable pulse energies as well as different positions of the focal spots posterior to the cornea were tested. Resulting lesions were evaluated by two-photon microscopy and histology to determine the best parameters for an exclusive detachment of corneal endothelial cells (CEC) with minimum collateral damage. The advantages of this method are the precise ablation of CEC, reduced collateral damage, and above all, the non-contact treatment.

Here, we present a protocol to detach corneal endothelial cells (CEC) from Descemet&#8217;s membrane (DM) using a neodymium:YAG (Nd:YAG) laser as an ex vivo disease model for bullous keratopathy (BK).

角膜内皮细胞损失非侵入性激光辅助实验模型的发展

Nd:YAG lasers have been used to perform noninvasive intraocular surgery, such as capsulotomy for several decades now. The incisive effect relies on the ...

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation of human liver transplantation due to the absence of arterial reconnection. Described here is a modified transplantation procedure that includes the incorporation of hepatic artery (HA) reconnection, leading to a marked improvement in transplant outcomes. With a mean anhepatic time of 12 min and 14 s, HA reconnection results in improved perfusion of the transplanted liver and an increase in long-term recipient survival from 37.5% to 88.2%. This protocol includes the use of 3D-printed cuffs and holders to connect the portal vein and infrahepatic inferior vena cava. It can be implemented for studying multiple aspects of liver transplantation, from immune response and infection to technical aspects of the procedure. By incorporating a simple and practical method for arterial reconnection using a microvascular technique, this modified rat OLT protocol closely mimics aspects of human liver transplantation and will serve as a valuable and clinically relevant research model.

The goal of this study is to modify the rat orthotopic liver transplant model to better represent human liver transplantation and improve recipient survival. The presented method reestablishes hepatic arterial inflow by connecting the donor liver&#39;s common hepatic artery to the recipient liver&#39;s proper hepatic artery.

矫形肝移植大鼠模型中动脉重联后并发症减少

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...

Orthotopic Kidney Auto-Transplantation in a Porcine Model Using 24 Hours Organ Preservation And Continuous Telemetry

In the present era of organ transplantation with critical organ shortage, various strategies are employed to expand the pool of available allografts for kidney transplantation (KT). Even though, the use of allografts from extended criteria donors (ECD) could partially ease the shortage of organ donors, ECD organs carry a potentially higher risk for inferior outcomes and postoperative complications. Dynamic organ preservation techniques, modulation of ischemia-reperfusion and preservation injury, and allograft therapies are in the spotlight of scientific interest in an effort to improve allograft utilization and patient outcomes in KT.
Preclinical animal experiments are playing an essential role in translational research, especially in the medical device and drug development. The major advantage of the porcine orthotopic auto-transplantation model over ex vivo or small animal studies lies within the surgical-anatomical and physiological similarities to the clinical setting. This allows the investigation of new therapeutic methods and techniques and ensures a facilitated clinical translation of the findings. This protocol provides a comprehensive and problem-oriented description of the porcine orthotopic kidney auto-transplantation model, using a preservation time of 24 hours and telemetry monitoring. The combination of sophisticated surgical techniques with highly standardized and state-of-the-art methods of anesthesia, animal housing, perioperative follow up, and monitoring ensure the reproducibility and success of this model.

Large animal models play an essential role in preclinical transplantation research. Due to its similarities to the clinical setup, the porcine model of orthotopic kidney auto-transplantation described in this article provides an excellent in vivo setting for the testing of organ preservation techniques and therapeutic interventions.

使用24小时器官保存和连续遥测的猪模型中正交肾自动移植

In the present era of organ transplantation with critical organ shortage, various strategies are employed to expand the pool of available allografts for ...

Intramyocardial Transplantation of MSC-Loading Injectable Hydrogels after Myocardial Infarction in a Murine Model

One of the major issues facing current cardiac stem cell therapies for preventing postinfarct heart failure is the low retention and survival rates of transplanted cells within the injured myocardium, limiting their therapeutic efficacy. Recently, the use of scaffolding biomaterials has gained attention for improving and maximizing stem cell therapy. The objective of this protocol is to introduce a simple and straightforward technique to transplant bone marrow-derived mesenchymal stem cells (MSCs) using injectable hydroxyphenyl propionic acid (GH) hydrogels; the hydrogels are favorable as a cell delivery platform for cardiac tissue engineering applications due to their ability to be cross-linked in situ and high biocompatibility. We present a simple method to fabricate MSC-loading GH hydrogels (MSC/hydrogels) and evaluate their survival and proliferation in three-dimensional (3D) in vitro culture. In addition, we demonstrate a technique for intramyocardial transplantation of MSC/hydrogels in mice, describing a surgical procedure to induce myocardial infarction (MI) via left anterior descending (LAD) coronary artery ligation and subsequent MSC/hydrogels transplantation.

Stem cell-based therapy has emerged as an efficient strategy to repair injured cardiac tissues after myocardial infarction. We provide an optimal in vivo application for stem cell transplantation using gelatin hydrogels that are able to be enzymatically cross-linked.

Murine 模型心肌梗塞后 MSC 加载注射水凝胶的心内移植

One of the major issues facing current cardiac stem cell therapies for preventing postinfarct heart failure is the low retention and survival rates of ...