作者感谢 e. Ferrantelli 和 g. Liappas 为他们的支持设置5/6 肾切除协议, R. Díaz 和P。马丁为协助与 ureic 氮气评估和 e. 何伟亚和 F. Núñez 为协助与老鼠关心。这项工作得到了 SAF2016-80648R 从 "Ministerio de Economía y Competitividad"/Fondo Europeo 区域 (收复与发展/MINECO) 向-卡夫和 PI 15/00598 的赠款的支持, 从菲德 de Fondo Investigaciones 基金, 到阿韦拉多·莫雷诺发言。

这里描述的所有方法都得到了分子生物学中心 Severo 奥乔亚 (西班牙马德里) 机构动物护理和使用委员会的批准。
注: 在研究开始时, C57BL/6J 雌性小鼠12岁至14周, 体重约20克。所有的动物都被安置在标准条件下, 并被给予食物和水的广告随意。每天检查健康状况。需要的材料, 如手套, 悬垂, 导管, 缝合和针头, 应该是无菌的。
1. 放置导管
注: 如果肾脏没有被切除, 他们仍然完全功能, 因此它不考虑尿毒症的作用, 因此允许研究 PDF 暴露在隔离。该手术包括将导管的远端极端引入腹膜腔, 并将进入端口置于动物的背部, 提供对其的访问。放置导管的程序如下:
<ol><li>将鼠标放置在感应腔内, 使用4% 异氟醚和氧气进行麻醉, 流速为0.4 升/分钟, 直至失去矫正反射。<ol>
<li>用与麻醉器具连接的 nosecone 管在100% 氧气中维持2% 异氟醚的动物, 其流量为0.3 升/分钟。通过评估肌肉的语调和对刺激的反应来确认正确的麻醉。</li>
		<li>检查整个过程中呼吸的速率和深度。在麻醉时使用眼部药膏, 防止干燥。最好的做法是在一个流动的内阁, 以确保在手术期间保持不育的条件。</li>
	</ol>
</li>
	<li>剃掉右侧面和动物的背部, 以便进行手术, 然后在干净的区域的入口口注射液体。将动物放置在手术台上左侧的侧位处, 用热系统确保其温度不会下降。</li>
	<li>用葡萄糖酸1% 洗必泰溶液对该区进行消毒。在身体右侧的皮肤上用钝剪刀做一个小切口 (0.5 厘米), 然后用剪刀从相邻的肌肉层仔细地将它分开, 这样, 动物背部的整个区域就可以很好地分离, 以后才能引入方便的访问端口。请参考图 1 , 查看遵循该过程所需的材料。</li>
	<li>通过肌肉层做一个直径约1毫米的小切口, 插入导管的尖端和第一个塑料环。造成的腹膜损伤极小。</li>
	<li>将腹膜壁紧紧地缝合在导管周围, 用5.0 或6.0 不可吸收缝合。一个塑料环, 然后位于腹腔内, 另一个在肌肉和皮肤之间。导管因此被固定, 以防止液体泄漏到皮下空间。</li>
	<li>将访问端口插入到皮下组织的小鼠尾部, 而不将其固定到皮肤的固定位置, 因为它可能会痒, 而且动物可能会擦伤并咬伤皮肤。</li>
	<li>用5.0 或6.0 不可吸收的缝线闭合皮肤的伤口。除去吸入麻醉, 让动物恢复知觉。不要让鼠标无人看管, 直到它恢复了足够的意识来维持胸骨卧床。当完全恢复时, 鼠标可以返回到其他动物的公司。 注: 实验可在术后4至7天后开始恢复。</li>
	<li>在手术当天, 在250毫升的饮用水中溶解3毫升布洛芬 (20 毫克/毫升), 提供镇痛作用。</li>
	<li>手术后, 检查动物每日的健康状况, 检查皮肤上没有红发区, 毛发或伤口。</li>
	<li>用一只手握住动物 (不麻醉), 用一只手掌抓住接入口, 用另一只手握针头, 注入液体。注射前用葡萄糖酸1% 洗必泰溶液对该区进行消毒。这是有趣的使用特殊的针头 (的针头), 这是斜面, 以部分而不是刺穿的硅胶隔膜的接入端口 (图 1A)。40天内每天两次注射足够观察腹膜改变 (图 3)。</li>
	<li>在完成实验后, 用二氧化碳窒息或颈椎脱位弄死老鼠。</li>
</ol>2. 执行5/6 肾切除术并放置导管
注: 为了更好地类似于 PD 患者的情况, 有可能执行5/6 肾切除术, 只允许残肾功能。在这种情况下, 应采取血清样本来分析尿素水平通过提取 250 ul 的血液通过面部静脉穿刺, 至少一天前开始手术, 在中间的治疗和当牺牲动物。最好的做法是在一个流动的内阁, 以确保在手术期间保持不育的条件。
<ol><li>在步骤1.1 中使用异氟醚麻醉小鼠。</li>
	<li>提供镇痛0.1 毫克/千克丁丙诺啡, 注射在动物的颈部和通过溶解3毫升的布洛芬 (20 毫克/毫升) 在250毫升的饮用水的前一天和一天的手术。</li>
	<li>剃掉分支和动物的背部, 以便进行手术, 然后在清洁区域的入口口注射液体。</li>
	<li>在皮肤、左侧、靠近肋骨的部位进行大约0.5 厘米的切口, 以便直接进入左肾。</li>
	<li>在肌肉中打开一个小切口, 从腹膜中取出左肾, 取出胶囊和肾上腺。为了更好地将肾脏放在腹膜腔外, 取出胶囊是必要的。</li>
	<li>用 cauterizer 烧伤并切开肾脏的极端 (见图 1所需材料)。</li>
	<li>将肾脏重新引入腹腔, 用不溶性的5.0 或6.0 缝线缝合肌肉和皮肤的伤口。</li>
	<li>后天, 完全取出右肾, 用同一个切口插入导管, 取出肾脏。再次, 麻醉鼠与异氟醚和皮下注射0.1 毫克/千克的丁丙诺啡在手术前。还可以在手术前的前一天和前一天将3毫升布洛芬 (20 毫克/毫升) 溶解在250毫升的饮用水中。</li>
	<li>在皮肤上切开0.5 厘米, 在剪刀的帮助下, 将动物背部的皮肤与肌肉分开, 打开进入端口所在的空间。</li>
	<li>在肌肉中 (大约 0.3-0.4 厘米) 进行切割, 将右肾从腹膜腔中取出。</li>
	<li>取出胶囊和肾上腺以更好地获得肾脏。结扎肾静脉、动脉和输尿管, 无吸收5.0 或6.0 缝合, 完全取出肾。</li>
	<li>缝合在腹膜肌肉的伤口, 引入导管的末端, 使肌肉必须保持在两个塑料环之间, 如前所述 (步骤 1.5)。</li>
	<li>将接入口引入皮下空间, 缝合皮肤, 如步骤1.6 和1.7 所述。 注: 小鼠应在这些手术中休息至少10天, 以确保腹腔肌肉的伤口完全愈合, 并在注射液体时不会漏入皮下空间。在步骤1.10 中注入液体。</li>
	<li>在完成实验后, 用二氧化碳窒息或颈椎脱位弄死动物。</li>
</ol>

作者没有什么可透露的。

首次发表的数据, 分析 PD 改变使用 "关闭系统" 技术进行了在 2009年29 。这个紧密的系统意味着整个设备位于体内, 流体通过一个入口端口注入针。通过导管输液的长期动物模型中最重要的技术问题是梗阻的发生。可能的选择是执行 omentectomy 或添加肝素到 pdf, 以减少腹膜粘连。然而, 网膜作为一个防御器官, 肝素除了抗凝作用外, 还能调节炎症细胞活性、血管生成、细胞外基质的合?...

肾脏功能与肾脏疾病
肾脏是参与稳态、血液过滤和荷尔蒙分泌的重要器官。有各种情况导致肾衰竭和随后发病的尿毒症, 这已被定义为组的系统性症状, 由于积累的血液中的废物, 由于肾脏功能紊乱保留1。此外, 由于肾功能衰竭时的自我平衡能力也受到影响, 可能发生体积超负荷的高血压, 这也是危险的, 因为它可能导致心力衰竭1。当肾脏功能能力小于 10%-15% 时, 患者必须接受下列治疗方案之一: 血液透析、腹膜透析 (PD) 或肾移植。
PD 是一个有趣的选择, 允许病人继续治疗, 从舒适的家庭或几乎任何地方, 从而避免需要频繁的医院访问和住宿。PD 技术通过渗透液 (腹膜透析液, PDF) 注入腹膜腔, 消除了人体2产生的小毒性分子和过量水。这种注入产生的渗透梯度, 以交换溶质和水之间的腹膜毛细管和 PDF, 一个过程称为超滤 (UF)。
腹膜透析引起的腹膜损伤
腹腔由一个膜 (PM) 所覆盖, 由一层间的间皮细胞组成, 它栖息在基质上, 它也很少有血管、成纤维细胞、巨噬细胞和其他体细胞群。不幸的是, 腹膜膜在 PD 治疗过程中总有一定的变化, 如细胞凋亡和间皮细胞的丧失, 内皮 (蒙脱石) 和内皮 (端 MT) 细胞的骨髓基质转移, 炎症细胞的招募和纤维细胞, 血管改变, 血管生成, 淋巴管形成和/或纤维化3,4,5,6,7,8,9。这些改变负责发展超滤功能衰竭10, 这妨碍了治疗的继续, 要求病人必须接受替代治疗生存 (血液透析或肾移植).因此, 对这些患者来说, 延缓或控制这些腹膜改变的发展是至关重要的。
据推测, 尿毒症单独可能引起炎症11, 但最重要的局部因素是 PDF bioincompatibility。大多数 pdf 使用葡萄糖作为渗透剂, 引起炎症。由于 PDF 存储时间和灭菌, 葡萄糖的降解过程, 新的产品从这个反应出现, 产生更多的炎症, 蒙脱土和凋亡12,13。此外, 由于灌注法还有机械损伤的可能性。所有这些因素, 持续不断, 可能产生持续和复发的炎症状态, 导致慢性炎症, 这推动膜恶化, 并最终, 超滤过的失败。如何减少或避免这种损害仍然是一个研究问题。
从人体标本到动物模型分析病变的发展
使用人体活检是一个限制因素, 由于难以获得组织样本。这些样品只能从因导管故障或移植而进行的手术中获得, 通常经过多年的 PD 治疗。这种方法有助于分析腹腔膜在 PDF 中所受的病理变化, 但不足以研究该过程的发展。另一种可能是分析透析出水排出的细胞, 但这仍然无法提供完整的方案。合并这两种技术只可能与动物模型。哺乳动物的腹膜结构相似, 因此有不同动物种类的模型。有几个基于绵羊的研究 (Rodela等。14和巴雷尔等。15) 及家兔16、17种机型;然而, 较小的动物是可取的, 因为他们更容易的住房和维护, 而且也更经济。使用鼠18,19,20,21,22,23,24提供了更短的治疗时间需要观察形态功能的改变。它代表了一个非常有用的模型来探索不同的问题, 如抗纤维化药物的作用, 例如 BMP-7 (骨形态 protein-7)25和 RAS (肾素-血管紧张系统) 靶向26,27,28。
然而, 小鼠模型已经成为一个理想的模型, 有许多好处比其他。最有趣的优势是利用基因修饰小鼠研究腹膜损伤的分子和细胞基础的可能性。事实上, 老鼠经常被用来分析许多疾病, 因为有许多不同的菌株与各种众所周知的遗传背景。其他好处包括住房所需的空间减少, 实验费用减少 (由于动物的尺寸较小), 易于处理, 试剂的可用性以及对不同品系的老鼠的可用信息数量的增加因为他们是最常用的动物在研究中。
使用植入装置的小鼠模型是最近建立的 pd29,30的模型, 并被证明模拟腹膜恶化的 pd 患者由于接触 pdf。这个模型合作了解的病理过程牵连31,32,33。此外, 它已被用来验证各种潜在的治疗, 以改善这种恶化使用免疫调节剂和抗炎药物和其他抗纤维化和抗血管生成药物, 如 COX-2 (cyclooxygenase-2) 抑制剂34, PPAR γ (过氧化物酶体核扩散激活受体γ) 激动剂35, 三苯氧胺36, Paricalcitol (维生素 D 受体活化剂, 调节免疫反应)37, 雷帕霉素38和 Nebivolol39。
用植入式导管研制小鼠模型
该模型的目的是尽可能类似于人类 pd 患者使用的技术, 允许对小动物的 pd 进行扩展治疗。到目前为止, 在小鼠中测试了三种将透析液注入腹膜的技术。第一个, 前腹壁盲穿刺, 是有争议的, 因为它可能招致的多重风险, 如腹膜损伤, 出血和盲目执行, 内脏穿刺。第二种技术是所谓的 "开放式永久性系统", 其中注入液体的装置放在身体外。这个过程是最相似的在人执行。然而, 它不允许长期实验的发展, 因为它可能增加传染的机会, 并且一般要求使用麻醉灌输 PDF, 可能干扰结果。第三种技术是 "封闭系统"。使用这种方法, 用于液体注入的整个装置位于动物体内。液体通过一根针注射在进入端口, 被放置在皮下。这一程序减少了腹膜感染和出血的风险以及麻醉的需要。
为了研究尿毒症在 PD 中的作用, 最近的小鼠模型也被 stablished40基于 PDF 输液模型的导管。该模型为小鼠进行肾切除术提供了一种新的方法, 从而降低了肾脏的功能。本文对 2015年40 Ferrantelli 的协议进行了修改. 这一新的协议允许导管植入术, 而肾切除, 减少伤口的长度, 在手术期间, 并方便进入肾脏。

<table><tbody><tr><td>Minute Mouse Port 4French with retention beads and cross holes</td><td>Access technologies</td><td>MMP-4S-061108A</td><td></td></tr><tr><td>Posi-Grip Huber point needles 25 ga. X 1/2&amp;acute;&amp;acute;&amp;nbsp;</td><td>Access technologies</td><td>PG25-500</td><td></td></tr><tr><td>High Temperature Cautery Kit</td><td>Bovie</td><td>18010-00</td><td></td></tr><tr><td>Forane</td><td>abbVie</td><td>880393.4 HO</td><td></td></tr><tr><td>non absorbable suture 6/0</td><td>Laboratorio Agar&amp;oacute;</td><td>6121</td><td></td></tr><tr><td>Scissors&amp;nbsp;</td><td>Fine Science Tools</td><td>14079-10</td><td></td></tr><tr><td>forceps</td><td>Fine Science Tools</td><td>11002-12</td><td></td></tr><tr><td>clamp</td><td>Fine Science Tools</td><td>13002-10</td><td></td></tr><tr><td>Buprenorphine 0,3 mg/ml</td><td></td><td></td><td>pharmaceutical product</td></tr><tr><td>cotton swabs</td><td></td><td></td><td>pharmaceutical product</td></tr><tr><td>Dalsy (Ibuprofen) 20mg/mL oral suspension</td><td>AbbVie&amp;nbsp;S.R.L.&amp;nbsp;</td><td></td><td>pharmaceutical product</td></tr></tbody></table>

surgical techniques for catheter placement and 5/6 nephrectomy in murine models of peritoneal dialysis

腹膜透析 (PD) 是一种肾脏替代疗法, 它与腹腔内高渗性液的管理和后恢复相一致, 用于排出水和有毒代谢物, 而功能不足的肾脏无法消除。不幸的是, 这个程序恶化了腹膜。组织损伤引发炎症愈合损伤。如果损伤持续, 炎症变得慢性, 可能导致纤维化, 这是许多疾病中常见的现象。在 PD 中, 慢性炎症和纤维化, 以及与这些相关的其他特定过程, 导致超滤能力的恶化, 这意味着技术的失败和随后的停止。使用人体样本提供有关这种恶化的信息, 但对获得活检有技术和伦理上的限制。动物模型是研究这种恶化的必要条件, 因为它们克服了这些缺点。
2008年开发了一种慢性小鼠输液模型, 它从广泛的基因修饰小鼠中受益, 开辟了研究相关机制的可能性。该模型采用了一种为小鼠设计的定制装置, 它由连接在动物背部的接入端口上的导管组成。这一程序避免了在长期实验中连续穿刺腹膜, 减少了注射引起的感染和炎症。由于该模型, 慢性 PD 液暴露引起的腹膜损伤具有明显的特点和调控作用。这项技术允许输液大量的液体, 并可用于研究其他疾病, 其中接种药物或其他物质在较长的时间内是必要的。
本文介绍了导管在小鼠中的手术放置方法。此外, 还解释了5/6 例肾切除术对 PD 患者肾功能不全状态的模拟。

图 1显示了遵循 "协议" 部分中描述的过程所需的所有材料。在这个例子中, 提交或不切除的老鼠 (每组8只动物) (图 2) 在40天内被暴露 (每天两次注射, 在两者之间至少等待2小时) 到两个不同 pdf 的混合物中, 通常用于临床实践:Extraneal (icodextrin pdf) 和 Dianeal (基于葡萄糖的 pdf)。stablished 为对照组 (n = 6)。
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Watch this Scientific Journal Video about Surgical Techniques for Catheter Placement and 5/6 Nephrectomy in Murine Models of Peritoneal Dialysis at JoVE.com

Surgical Techniques for Catheter Placement and 5/6 Nephrectomy in Murine Models of Peritoneal Dialysis

小鼠腹膜透析模型导管置入及5/6 切除术的手术技术

这篇文章显示了手术放置在小鼠腹腔导管连接到一个接入端口, 位于动物的背部。此外, 它还解释了5/6 例肾切除术与 PD 患者尿毒状态相似的过程。

Peritoneal dialysis (PD) is a renal replacement therapy consistent on the administration and posterior recovery of a hyperosmotic fluid in the peritoneal ...

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15.1K 次观看.  Spanish National Research Council. 这篇文章显示了手术放置在小鼠腹腔导管连接到一个接入端口, 位于动物的背部。此外, 它还解释了5/6 例肾切除术与 PD 患者尿毒状态相似的过程。

视频: Surgical Techniques for Catheter Placement and 5/6 Nephrectomy in Murine Models of Peritoneal Dialysis

Isolation of Glomeruli and In Vivo Labeling of Glomerular Cell Surface Proteins

Proteinuria results from the disruption of the glomerular filter that is composed of the fenestrated endothelium, glomerular basement membrane, and podocytes with their slit diaphragms. The delicate structure of the glomerular filter, especially the slit diaphragm, relies on the interplay of diverse cell surface proteins. Studying these cell surface proteins has so far been limited to in vitro studies or histologic analysis. Here, we present a murine in vivo biotinylation labeling method, which enables the study of glomerular cell surface proteins under physiologic and pathophysiologic conditions. This protocol contains information on how to perfuse mouse kidneys, isolate glomeruli, and perform endogenous immunoprecipitation of a protein of interest. Semi-quantitation of glomerular cell surface abundance is readily available with this novel method, and all proteins accessible to biotin perfusion and immunoprecipitation can be studied. In addition, isolation of glomeruli with or without biotinylation enables further analysis of glomerular RNA and protein as well as primary glomerular cell culture (i.e., primary podocyte cell culture).

Isolation of Glomeruli and In Vivo Labeling of Glomerular Cell Surface Proteins

Here we present a protocol for murine in vivo labeling of glomerular cell surface proteins with biotin. This protocol contains information on how to perfuse mouse kidneys, isolate glomeruli, and perform endogenous immunoprecipitation of the protein of interest.

肾小球的分离及肾小球细胞表面蛋白的活体标记

Proteinuria results from the disruption of the glomerular filter that is composed of the fenestrated endothelium, glomerular basement membrane, and ...

科研

生物化学

Modeling Spontaneous Metastatic Renal Cell Carcinoma (mRCC) in Mice Following Nephrectomy

One of the key challenges to improved testing of new experimental therapeutics in renal cell carcinoma (RCC) is the development of models that faithfully recapitulate early- and late-stage metastatic disease progression. Typical tumor implantation models utilize ectopic or orthotopic primary tumor implantation, but few include systemic spontaneous metastatic disease that mimics the clinical setting. This protocol describes the key steps to develop RCC disease progression stages similar to patients. First, it uses a highly metastatic mouse tumor cell line in a syngeneic model to show orthotopic tumor cell implantation. Methods include superficial and internal implantation into the sub-capsular space with cells combined with matrigel to prevent leakage and early spread. Next it describes the procedures for excision of tumor-bearing kidney (nephrectomy), with critical pre- and post- surgical mouse care. Finally, it outlines the steps necessary to monitor and assess micro-and macro-metastatic disease progression, including bioluminescent imaging as well provides a detailed visual necropsy guide to score systemic disease distribution. The goal of this protocol description is to facilitate the widespread use of clinically relevant metastatic RCC models to improve the predictive value of future therapeutic testing.&#160;

Modeling Spontaneous Metastatic Renal Cell Carcinoma mRCC in Mice Following Nephrectomy

Models of spontaneous metastatic renal cell carcinoma (RCC) disease progression can be used for evaluating treatments in a clinically relevant setting. This protocol demonstrates different procedures for orthotopic kidney tumor cell implantation, proper nephrectomy, and finally outlines a necropsy guide for visual and bioluminescent scoring of metastatic burden and localization.

在小鼠模型自发转移性肾细胞癌（MRCC）肾切除

One of the key challenges to improved testing of new experimental therapeutics in renal cell carcinoma (RCC) is the development of models that faithfully ...

医学

Murine Kidney Transplant Technique

The first mouse kidney transplant technique was published in 19731 by the Russell laboratory. 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 using the donor aorta to form the arterial anastomosis instead of the renal artery was developed and reported in 1993 by Kalina and Mottram 2 with a further advancement coming from the same laboratory in 1999 3. While one can become proficient in this model, a search of the literature reveals that many labs still experience a high proportion of graft loss due to arterial thrombosis. We describe here a technique that was devised in our laboratory that vastly reduces the arterial thrombus reported by others 4,5. This is achieved by forming a heel-and-toe cuff of the donor infra-renal aorta that facilitates a larger anastomosis and straighter blood flow into the kidney.

The goal of this manuscript is to describe the steps required to perform a kidney transplant in a mouse, paying particular attention to the details of the arterial anastomosis.

小鼠肾脏移植技术

The first mouse kidney transplant technique was published in 19731 by the Russell laboratory. Although it took some years for other labs to become ...

Microdissection of Primary Renal Tissue Segments and Incorporation with Novel Scaffold-free Construct Technology

Kidney transplantation is now a mainstream therapy for end-stage renal disease. However, with approximately 96,000 people on the waiting list and only one-fourth of these patients achieving transplantation, there is a dire need for alternatives for those with failing organs. In order to decrease the harmful consequences of dialysis along with the overall healthcare costs it incurs, active investigation is ongoing in search of alternative solutions to organ transplantation. Implantable tissue-engineered renal cellular constructs are one such feasible approach to replacing lost renal functionality. Here, described for the first time, is the microdissection of murine kidneys for isolation of living corticomedullary renal segments. These segments are capable of rapid incorporation within scaffold-free endothelial-fibroblast constructs which may enable rapid connection with host vasculature once implanted. Adult mouse kidneys were procured from living donors, followed by stereoscope microdissection to obtain renal segments 200 - 300 &#181;m in diameter. Multiple renal constructs were fabricated using primary renal segments harvested from only one kidney. This method demonstrates a procedure which could salvage functional renal tissue from organs that would otherwise be discarded.

Tissue-engineered renal constructs provide a solution for the organ shortage and deleterious effects of dialysis. Here, we describe a protocol to micro dissect murine kidneys for isolation of cortico-medullary segments. These segments are implanted into scaffold-free cellular constructs, forming renal organoids.

原发性肾组织段的显微解剖与新型无支架结构技术的结合

Kidney transplantation is now a mainstream therapy for end-stage renal disease. However, with approximately 96,000 people on the waiting list and only ...

生物工程

A Modified Surgical Technique for Kidney Transplantation in Mice

Kidney transplantation in mice is a complicated and challenging surgery procedure. There are very few publications demonstrating the key steps of this operation. Therefore, this article introduces the technique and points out the surgical caveats associated with this operation. In addition, important modifications in comparison to the conventional procedure are demonstrated. Firstly, a patch of the abdominal aorta is cut and prepared so that the proximal bifurcations of the renal artery, including the ureteral artery are transected together with the donor kidney en bloc. This reduces the risk of a ureter necrosis and avoids the development of a urinary tract occlusion. Secondly, a new method of the vascular anastomosis is demonstrated that allows the operator to flexibly increase or decrease the size of the anastomosis after renal transplant reperfusion has already been initiated. This avoids the development of vessel strictures and intraabdominal bleeding. Thirdly, a technique that enables the anastomosis of the delicate donor ureter and the recipient bladder that does not cause a trauma is shown. Adopting this protocol can shorten the operation time and reduces the damage to the recipient's bladder, thereby significantly increasing the operation success rate for the recipient mice.

This protocol presents a new surgical technique of mouse kidney transplantation focusing on a modified arterial anastomosis strategy. A vascular suture technique including a simple and safer ureter-bladder anastomosis method is also presented. These modifications shorten the operation time and improve the success rate of the mouse kidney transplantation procedure.

一种用于小鼠肾移植的改良手术技术

Kidney transplantation in mice is a complicated and challenging surgery procedure. There are very few publications demonstrating the key steps of this ...

免疫与感染

A Retrograde Implantation Approach for Peritoneal Dialysis Catheter Placement in Mice

Murine models are employed to probe various aspects of peritoneal dialysis (PD), such as peritoneal inflammation and fibrosis. These events drive peritoneal membrane failure in humans, which remains an area of intense investigation due to its profound clinical implications in managing patients with end-stage kidney disease (ESKD). Despite the clinical importance of PD and its related complications, current experimental murine models suffer from key technical challenges that compromise the models&#39; performance. These include PD catheter migration and kinking and usually warrant earlier catheter removal. These limitations also drive the need for a greater number of animals to complete a study. Addressing these drawbacks, this study introduces technical improvements and surgical nuances to prevent commonly observed PD catheter complications in a murine model. Moreover, this modified model is validated by inducing peritoneal inflammation and fibrosis using lipopolysaccharide injections. In essence, this paper describes an improved method to create an experimental model of PD.

This article describes modifications of a procedure to implant a peritoneal dialysis catheter in a murine model to avoid major technical issues observed with the conventional techniques.

一种小鼠腹膜透析导管放置的逆行植入方法

Murine models are employed to probe various aspects of peritoneal dialysis (PD), such as peritoneal inflammation and fibrosis. These events drive ...

A Murine Model of Hemodialysis Access-Related Hand Dysfunction

Chronic kidney disease is a major public health problem, and the prevalence of end-stage renal disease (ESRD) requiring chronic renal replacement therapies such as hemodialysis continues to increase. Autogenous arteriovenous fistula (AVF) placement remains a primary vascular access option for ESRD patients. Unfortunately, approximately half of the hemodialysis patients experience dialysis access-related hand dysfunction (ARHD), ranging from subtle paresthesia to digital gangrene. Notably, the underlying biologic drivers responsible for ARHD are poorly understood, and no adequate animal model exists to elucidate the mechanisms and/or develop novel therapeutics for the prevention/treatment of ARHD. Herein, we describe a new mouse model in which an AVF is created between the left common iliac artery and vein, thereby facilitating the assessment of limb pathophysiology. The microsurgery includes vessel isolation, longitudinal venotomy, creation of arteriovenous anastomosis, and venous reconstruction. Sham surgeries include all the critical steps except for AVF creation. Iliac AVF placement results in clinically relevant alterations in central hemodynamics, peripheral ischemia, and impairments in hindlimb neuromotor performance. This novel preclinical AVF model provides a useful platform that recapitulates common neuromotor perturbations reported by hemodialysis patients, allowing researchers to investigate the mechanisms of ARHD pathophysiology and test potential therapeutics.

This protocol details the surgical steps of murine common iliac arteriovenous fistula creation. We developed this model to study hemodialysis access-related limb pathophysiology.

血液透析通路相关手功能障碍的小鼠模型

Chronic kidney disease is a major public health problem, and the prevalence of end-stage renal disease (ESRD) requiring chronic renal replacement ...

A Mouse 5/6th Nephrectomy Model That Induces Experimental Uremic Cardiomyopathy

Chronic kidney disease (CKD) is a great risk factor for cardiovascular disease events and mortality, and progressively develops to the clinical phenotype called "uremic cardiomyopathy". We describe here an experimental CKD mouse model, named 5/6th partial nephrectomy (PNx) with pole ligation, which developed uremic cardiomyopathy at four weeks post-surgery. This PNx model was performed by a two-step surgery. In step-one surgery, both poles of the left kidney were ligated. In step-two surgery, which was performed 7 days after the step-one surgery, the right kidney was removed. For the sham surgery, the same surgery procedures were performed but without pole ligation of the left kidney or removal of the right kidney. The surgical procedures are easier and less time-consuming, compared to other methods. However, the remnant functional renal mass is not as easily controlled as the renal artery ligation. Four weeks after surgery, in comparison with the sham-operated mice, the PNx mice developed impaired renal function, anemia, cardiac hypertrophy, cardiac fibrosis, and decreased heart systolic and diastolic function.

A Mouse 5/6th Nephrectomy Model That Induces Experimental Uremic Cardiomyopathy

This manuscript provides a detailed two-step surgical procedure to perform mouse 5/6th partial nephrectomy (PNx) with pole ligation. Four weeks after surgery, in comparison with sham-operated mice, the PNx mice developed impaired renal function, anemia, cardiac hypertrophy, cardiac fibrosis, and decreased heart systolic and diastolic function.

一种诱导实验性尿毒症心肌病的小鼠 5/6th肾切除模型

Chronic kidney disease (CKD) is a great risk factor for cardiovascular disease events and mortality, and progressively develops to the clinical phenotype ...

生物学

<meta charset="utf8"></head><body>大鼠标准化 5/6 肾切除术治疗肾功能不全和灌注不良

5/6 Nephrectomy Using Sharp Bipolectomy Via Midline Laparotomy in Rats

Chronic kidney disease (CKD) affects over 10% of the global population, amounting to more than 800 million individuals worldwide. The advances in the treatment of CKD have had a significant impact on patient outcomes. While in the past, CKD was often considered a death sentence, with many patients succumbing to the complications of end-stage renal disease, it is now increasingly being managed as a chronic condition with the availability of dialysis and kidney transplantation, as well as new pharmaceutical developments such as SGLT2 inhibitors or nonsteroidal mineralocorticoid receptor antagonists. 
Yet, there continues to be a growing demand for further exploration into the pathophysiological processes and potential therapeutic interventions. Reliable biological models play a crucial role in facilitating this research. Given the multifaceted nature of renal disease, which encompasses not only cell biology but also vascular microanatomy and endocrine signaling, an appropriate model must possess a level of biological complexity that only an animal model can offer, rendering rodents an obvious choice.
This manuscript, therefore, provides an intricate, systematic protocol for surgically reducing renal parenchyma through midline laparotomy and combined total and partial nephrectomy in rats for survival as well as non-survival applications. It emphasizes the critical role of precise surgical techniques in ensuring consistent and reliable outcomes. Prime examples of potential applications for this model include biomolecular and pharmaceutical studies as well as the development of innovative intraoperative imaging modalities, such as hyperspectral imaging, to objectively visualize and differentiate renal malperfusion.

<meta charset="utf8"></head><body>5/6 大鼠使用中线剖腹手术进行 Sharp Bipolectomy 的肾切除术

This manuscript introduces a standardized protocol for a 5/6 nephrectomy by sharp bipolectomy via midline laparotomy in a rat model, aiming to induce a state of renal insufficiency via renal parenchyma reduction with a great degree of methodical precision and low risk of technical error.

5/6 大鼠使用中线剖腹手术进行 Sharp Bipolectomy 的肾切除术

Chronic kidney disease (CKD) affects over 10% of the global population, amounting to more than 800 million individuals worldwide. The advances in the ...

小鼠腹膜透析模型导管置入及5/6 切除术的手术技术

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此视频中的章节

肾小球的分离及肾小球细胞表面蛋白的活体标记

在小鼠模型自发转移性肾细胞癌（MRCC）肾切除

小鼠肾脏移植技术

原发性肾组织段的显微解剖与新型无支架结构技术的结合

一种用于小鼠肾移植的改良手术技术

一种小鼠腹膜透析导管放置的逆行植入方法

血液透析通路相关手功能障碍的小鼠模型

一种诱导实验性尿毒症心肌病的小鼠 5/6^th肾切除模型

5/6 大鼠使用中线剖腹手术进行 Sharp Bipolectomy 的肾切除术

5/6 大鼠使用中线剖腹手术进行 Sharp Bipolectomy 的肾切除术