我们要感谢Reizes实验室的成员在编写手稿期间所做的批判性审查和见解，以及Lerner研究所的成像和组织学核心在数据收集和数据分析方面给予的帮助。这项工作通过克利夫兰诊所的研究计划委员会提供内部赠款资金，并通过生殖调查协会和拜耳获得外部赠款。Reizes实验室的研究也通过维洛萨诺自行车治疗，妇科癌症研究中心，并通过劳拉J.福加蒂赋予子宫癌研究主席资助。克利夫兰诊所拥有图1和图2的版权许可。

注:在这项研究中使用动物得到了克利夫兰诊所勒纳研究所的机构动物护理和使用委员会（IACUC）的批准。所有公开的动物护理和使用标准都是根据国家卫生研究院的指导方针执行的。此过程采用无菌技术。佩特里菜是无菌的。使用的PBS/盐水是无菌的。用于尸检和组织解剖的手术器械通过高压灭菌器进行灭菌。我们在动物病例之间的仪器上使用 70% 的 EtOH（如果每节会话超过一次），以减少污染。
1.准备捐赠者和受体小鼠（见图1）
<ol><li>确保获得适当的批准，以便与实验室动物合作。</li>
	<li>确定鼠标的应变。在这些实验中，利用了具有C57BL/6J背景的野生型和突变小鼠，但使用类似模型的研究人员报告了与其他小鼠株（如BALB/c小鼠10）的成功。此外，捐赠者和/或受体小鼠可以使用记者系统，如GFP或RFP小鼠（见图2和图3）。</li>
	<li>对于子宫内膜组织移植的时间，确保供体小鼠在注射戈纳多特罗平时年龄在22-24天之间。这确保了他们在生殖上天真，例如，还没有开始可怕的骑自行车。</li>
	<li>确保受体小鼠在2至4个月大之间生殖完整（无前卵巢切除术）。虽然不需要，但建议定期将男性小鼠笼子里的尿液浸泡床上用品放在接受者的笼子里，以方便持续的腹膜循环，并在子宫内膜组织移植前72小时再次放置。 注:这不能确保接受者尿浸的床上用品的放置高度取决于男性床上用品的使用量，并且结果参差不齐。如果需要异质周期的同步，连续三次100 ng/100 μL雌二醇的皮下注射将使所有动物进入最恶劣的阶段。虽然我们发现基于卵清循环相的病变诱导没有差异，但其他组发现周期阶段很重要。</li>
	<li>皮下注射怀孕母马血清果纳多特罗平（PMSG，2 IU稀释成200μL）到捐赠小鼠皮下使用细针（25-27G推荐）。不要给受体小鼠PMSG，因为它会触发排卵和随后的高黄体酮环境，这是不太接受子宫内膜异位症的形成。 注:先前的小鼠模型已经利用PMSG或雌激素刺激子宫内膜增殖在捐赠小鼠子宫内膜收获23之前。PMSG 的建议原因如下:PMSG 的半衰岁为 40 小时，而 17-β-雌二醇的半衰岁仅为 2 小时。在采购子宫内膜组织之前，在捐赠者中采用单次皮下注射PMSG，可持续持续接触腺激素刺激，从而刺激内源性雌激素。外源性雌激素的许多制剂需要多次注射才能充分给子宫内膜调高。</li>
	<li>PMSG 注射后，安排尸检、采购和移植到接受者 38-42 小时之间。在注射后，子宫内膜组织收获的时间，以确保排卵前收集组织，排卵后发生于42小时。排卵产生高黄体酮 （P4） 环境， 这将减少病变的建立.</li>
</ol>2. 采购供体小鼠子宫内膜组织
<ol><li>捐赠小鼠安乐死后（使用CO2室后颈椎错位），用70%乙醇溶液喷洒腹部:这有助于减少从皮肤植物和脱落的头发采购的组织污染。用解剖剪刀，在中线通过皮肤和皮下组织进行浅横向剪裁。然后抓住皮肤切口的每一侧，用钝牵引力打开腹部。</li>
	<li>识别子宫。切除子宫之前，修剪掉相邻的结缔组织。将每个子宫角切开，就在各自的输卵管下方，然后转入子宫颈，切除整个子宫群。</li>
	<li>从腹腔切除后，仔细检查子宫，并去除任何额外的外周脂肪或结缔组织。将子宫放在一滴冷的PBS上，放在培养皿上。确定并记录整个子宫的组合质量。</li>
	<li>从子宫内切除每个角，使转角尽可能接近子宫，从而最大限度地延长每个角的长度。使用解剖显微镜的辅助，将解剖剪刀的一刀片放在第一角的流明内，然后沿着管子的主要轴线切割。小心地打开管子，记住哪一边是血清，哪一边是上皮。</li>
	<li>将 500 cc 的盐水或 PBS 放在新的培养皿上;液体会由于表面张力而保持在一起。</li>
	<li>然后，以统一的方式进行子宫分裂。与许多较小的病变相比，更大的病变要少。首先，通过抓住子宫内膜层并剥去其去皮，将上皮从肌膜中分离出来。<ol>
<li>或者，只需在不分离肌膜的情况下分割组织（只要上皮侧完全暴露），但保留肌膜会降低该模型与人类疾病的生理相关性。确保碎片尽可能大，但足够小，足以通过18G针;（建议为 1 mm x 1 mm）。</li>
		<li>从第一角收集这些 1 mm x 1 mm 片段中的 10-12 个（大致相当于 C57BL/6J 小鼠的 40 毫克组织）。将收集的碎片放入液体收集中。</li>
	</ol>
</li>
	<li>执行相同的步骤为另一个子宫角共约24个片段每只小鼠。记录碎片总数。</li>
</ol>3. 将组织片段腹膜注射到受体小鼠体内
<ol><li>使用 1 cc 注射器的钝端吸气悬挂的 1 mm x 1 mm 碎片;总容积应为 1 mL。</li>
	<li>将 18 G 针头连接到整个注射器上，并将液体加载到针中。考虑模拟注射回培养皿，以确保所有的组织将通过针。</li>
	<li>取收件鼠标。在IP注射之前或之后，获得接受者小鼠的阴道涂片，以获得阴道循环文档（10 μL盐水使用阴道孔的灯泡注射器，并用盖唇镀在玻璃滑梯上）。</li>
	<li>用注射器以45度角对片段进行腹内注射，注意不要皮下注射。</li>
	<li>如果注射后碎片仍然存在，请将额外的 200 μL 液体抽入注射器，以确保所有片段均在腹中成功注射。</li>
	<li>一旦保证没有出血或并发症，将受体小鼠放回笼子里，并喂养正常饮食。</li>
</ol>4. 子宫内膜病变的收获
<ol><li>移植后约21天对受体小鼠实施安乐死。 注:根据经验和使用类似模型的合作者的讨论，最大病变大小和数量发生在移植后约3周:3周后，病变开始缩小。使用国际自然保护联盟批准的安乐死方法。</li>
	<li>安乐死和颈椎错位后，用70%的乙醇喷洒动物腹部，用解剖剪刀将皮肤切成表面。用剪刀将皮肤和皮下空间切开，直截了当地打开腹部。</li>
	<li>在进行进一步解剖之前，对毛病变进行全面调查，用卡钳测量大小，并记录其解剖区域（见下文）。</li>
	<li>完整收集三个不同的解剖区域（无论病变是否可被严重欣赏）。如果病变在其他区域（如肠道）看到，这些病变应被忽略，除非病变穿过下面的三个区域之一，否则不应收集这些病变。<ol>
<li>A = 腹壁/腹膜（只要样品之间一致，都可以被扁平成盒式磁带或卷起）。</li>
		<li>B = 胰腺和中性脂肪。</li>
		<li>C = 副宫结缔组织和脂肪（围绕子宫的白色闪闪发光的组织，但不涉及膀胱以外的任何器官;注意不要将膀胱误认为是病变）。</li>
	</ol>
</li>
	<li>将每个解剖区域放在盒式磁带中，并适当标记，并根据组织切除的实验室协议将其放入正式和处理中。</li>
	<li>在两个均匀的深度下，每个组织区域在两个幻灯片（D1 和 D2）中分块正式块。</li>
</ol>5. 子宫内膜病变评分
<ol><li>扫描（放大 40 倍）和存档幻灯片。</li>
	<li>使用数字幻灯片阅读软件，定义子宫内膜病变边缘之间的最长距离 （X） - 无论边缘由腺体还是频闪组成，并标记它。连续病变由被频闪包围的腺体定义:该线不一定只穿过子宫内膜组织（如确定子宫内膜异位症不规则形状或起伏焦点的终点），但两个终点需要通过连续频闪和/或腺体连接（见图4）。</li>
	<li>使第二行 （Y） 跨越第一行 90 度，第二行的长度按照上述规则确定。</li>
	<li>如果遇到多个非连续病变，则给每个病变自己的 X 和 Y 测量。</li>
	<li>将每张幻灯片的最终分数计算为每个病变区域 （X*Y） 的汇总。</li>
	<li>将两张幻灯片（D1 vs D2）中较大的分数作为该区域（A、B、C）的最终分数。</li>
	<li>将每个区域的分数总计为该动物提供最终的微观分数。</li>
</ol>

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作者没有利益冲突可以披露。

我们的研究表明，子宫内膜异位症可以在小鼠中可靠诱导，而无需使用卵巢切除术和/或生存手术，并且通过腹部的标准化调查和组织分析可以识别和量化异位子宫内膜病变。
许多子宫内膜异位症的研究利用手术诱发的子宫内膜异位症，其中供体子宫内膜被缝合到位到肠道，腹壁，或其他腹腔内位置<sup cla...

子宫内膜异位症是女性生殖道的一种神秘疾病，给1、2岁女性带来沉重的经济和健康负担。子宫内膜异位症的病因尚未完全了解，并提出了多种解释，包括宇宙代谢、胚胎穆勒安息、骨髓衍生祖细胞的招募和逆行月经3。虽然这些拟议机制的多个方面可能涉及，而且没有单一的解释可以解释该疾病的所有形式，但子宫内膜异位症发病机制的主要模式是逆行月经。逆行月经是月经污水通过输卵管进入腹腔:据估计，90%的月经妇女定期接受逆行月经4，5。鉴于这种常见的月经逆行现象，为什么子宫内膜异位症只在一部分妇女中发展尚不清楚。为了更好地了解这种疾病的病因，直接的人类研究是不可行的，动物研究是有道理的。
子宫内膜异位症是治疗和研究的挑战。该病的流行率尚不清楚，但估计为10%1。虽然一些先进的子宫内膜异位症可以通过非侵入性成像准确识别，但只有通过对手术获得的活检标本进行组织病理分析才能得出明确的诊断:在视觉上看起来是病变的病变，实际上可能是纤维化或疤痕从其他原因6。疾病的严重程度和程度与症状7无关。
子宫内膜异位症病变由异质细胞类型和群体组成，这些细胞类型和群体在微环境内以复杂的方式相互作用，因此限制了细胞模型8、9的用处。在体内模型存在，但这些有内在的挑战和局限性10，11，12。灵长类动物模型是理想的，但往往不可行13，14，15。很少有非灵长类哺乳动物月经并自发地发展为子宫内膜异位症。子宫内膜异位症的啮齿动物模型存在，但每个模型都有限制17。其中许多模型需要生存手术缝合或植入子宫内膜组织到捐赠者接受壁或肠道，增加了技术复杂性，需要麻醉，并混淆免疫因素从手术本身18，19，20。此外，许多模型需要卵巢切除术和雌激素补充:在增加病变产量的同时，这增加了时间、费用和额外的生存手术。腹膜（IP）注射模型不需要麻醉或生存手术，这些模型逻辑上模拟逆行月经比缝合模型21，22，23更好。然而，由于注射后子宫内膜片段的随机分散，以及因此在病变识别和测量方面更加偏差，大多数 IP 模型在病变位置的变异性更大。
在这里，我们提出了子宫内膜异位症的穆林模型，将现有子宫内膜异位症模型的几个特征整合到一个新颖、简化和高效的系统中，该系统依赖于微观量化来代替主观分级。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Supplies for injecting PMSG into donor mouse</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1 mL Tuberculin syringe with 27G needle</td>
			<td>Fisher Scientific</td>
			<td>14-826-87</td>
			<td></td>
		</tr>
		<tr>
			<td>Pregnant mare serum gonadotropin</td>
			<td>Sigma-Aldrich</td>
			<td>9002-70-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Supplies for necropsy of donor mouse and tissue processing</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>6&#8221; serrated forceps, curved tip</td>
			<td>Electron Microscopy Sciences</td>
			<td>72993-6C</td>
			<td></td>
		</tr>
		<tr>
			<td>70% ethanol solution</td>
			<td>Pharmco</td>
			<td>33000HPLCCS4L</td>
			<td>70% solution dilute ethyl acetate 200 proof</td>
		</tr>
		<tr>
			<td>Analytical balance</td>
			<td>Mettler Toledo</td>
			<td>ME54TE</td>
			<td></td>
		</tr>
		<tr>
			<td>Carbon dioxide</td>
			<td>TriGas Supplier</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting tray</td>
			<td>Fisher Scientific</td>
			<td>S14000</td>
			<td></td>
		</tr>
		<tr>
			<td>No. 10 disposable scalpel</td>
			<td>Fisher Scientific</td>
			<td>NC9999403</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors, curved</td>
			<td>Electron Microscopy Sciences</td>
			<td>72941</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors, straight</td>
			<td>Electron Microscopy Sciences</td>
			<td>72940</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereo microscope</td>
			<td>Leica Microsystems</td>
			<td>Leica SE 4</td>
			<td>For tissue dissection</td>
		</tr>
		<tr>
			<td>Sterile phosphate buffered saline (PBS)</td>
			<td>Institutional core facility supplies</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical instrument sterilization tray</td>
			<td>Electron Microscopy Sciences</td>
			<td>66112-02</td>
			<td></td>
		</tr>
		<tr>
			<td>Tissue culture dishes</td>
			<td>Fisher Scientific</td>
			<td>08-772E</td>
			<td></td>
		</tr>
		<tr>
			<td>Weighing dishes</td>
			<td>Fisher Scientific</td>
			<td>02-202-103</td>
			<td></td>
		</tr>
		<tr>
			<td>Supplies for injecting into recipient mouse</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1 cc syringe</td>
			<td>BD Biosciences</td>
			<td>301025</td>
			<td></td>
		</tr>
		<tr>
			<td>18 G needle</td>
			<td>Fisher Scientific</td>
			<td>148265d</td>
			<td></td>
		</tr>
		<tr>
			<td>200 uL pipette tip</td>
			<td>Fisher Scientific</td>
			<td>02-707-422</td>
			<td></td>
		</tr>
		<tr>
			<td>Double distilled water</td>
			<td>Institutional core facility supplies</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Latex bulb</td>
			<td>Fisher Scientific</td>
			<td>03-448-21</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro cover glass slip</td>
			<td>VWR</td>
			<td>48366-067</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscope slide</td>
			<td>Fisher Scientific</td>
			<td>12-544-7</td>
			<td></td>
		</tr>
		<tr>
			<td>Standard light microscope</td>
			<td>Leica Microsystems</td>
			<td>DM IL</td>
			<td>For evaluating vaginal cytology smears</td>
		</tr>
		<tr>
			<td>Supplies for harvesting tissue from recipient mouse</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>10% Buffered formalin</td>
			<td>Fisher Scientific</td>
			<td>SF100-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Biopsy foam pads</td>
			<td>Fisher Scientific</td>
			<td>22-038-222</td>
			<td></td>
		</tr>
		<tr>
			<td>Precision Digital Calipers</td>
			<td>Electron Microscopy Sciences</td>
			<td>62065-40</td>
			<td></td>
		</tr>
		<tr>
			<td>Processing/embedding cassettes</td>
			<td>Fisher Scientific</td>
			<td>22-272416</td>
			<td></td>
		</tr>
	</tbody>
</table>

a syngeneic murine model of endometriosis using naturally cycling mice

子宫内膜异位症是骨盆疼痛和不孕症的主要原因。它是由子宫外部位的子宫内膜组织的存在来定义的。子宫内膜异位症新疗法和诊断工具的开发受到限制，部分原因是在研究这种疾病方面面临挑战。除灵长类动物外，很少有哺乳动物月经，也没有一种会出现自发性子宫内膜异位症。啮齿动物模型很受欢迎，但需要人工诱导子宫内膜异位症，许多模型利用免疫功能低下的小鼠或手术诱发的疾病。最近，人们更加关注涉及腹内注射的模型。我们提出了子宫内膜异位症的模糊模型，将现有子宫内膜异位症模型的几个特征整合到一个新的简化系统中，该系统依靠微观量化代替主观分级。在这个模型中，我们执行激素刺激的捐赠小鼠，腹腔注射，系统的腹部调查和组织收获，和组织定量，可以在坏死后的任何时间进行和验证。这种模式需要最少的资源和培训:不需要实验室技术人员在穆林生存手术或确定子宫内膜病变方面的专业知识;可用于免疫功能低下、免疫能力和/或突变小鼠:并可靠地创建与人类子宫内膜疾病在组织学上一致的子宫内膜病变。

为了概念实验的初步证明，从RFP小鼠的供体子宫内膜被注射到野生型受体小鼠。H&E染色揭示了子宫内膜异位症病变经典结构的组织病理学确认（图3A）。荧光显微镜证实，观察到的病变来自供体（图3B）。
第二个实验使用10个野生类型C57BL/6J捐赠者和10个接受者进行。另有5名接受者接受了虚假治疗（注射了PBS，而不是子宫内膜片段）...

Watch this Scientific Journal Video about A Syngeneic Murine Model of Endometriosis using Naturally Cycling Mice at JoVE.com

A Syngeneic Murine Model of Endometriosis using Naturally Cycling Mice

许多啮齿动物子宫内膜异位症模型受技术复杂性、可重复性和/或免疫功能低下的动物或特殊记者小鼠的需求的限制。我们使用任何实验鼠使用独立可验证的客观评分系统，无需进行卵巢切除术或生存手术，即可提供简化的病变诱导系统。

使用自然循环小鼠的子宫内膜异位症的合成穆林模型

Endometriosis is a leading cause of pelvic pain and infertility. It is defined by the presence of endometrial tissue in extrauterine locations. The ...

a-syngeneic-murine-model-of-endometriosis-using-naturally-cycling-mice

Research

JoVE Journal

Medicine

4.9K 次观看.  Cleveland Clinic. 许多啮齿动物子宫内膜异位症模型受技术复杂性、可重复性和/或免疫功能低下的动物或特殊记者小鼠的需求的限制。我们使用任何实验鼠使用独立可验证的客观评分系统，无需进行卵巢切除术或生存手术，即可提供简化的病变诱导系统。

视频: A Syngeneic Murine Model of Endometriosis using Naturally Cycling Mice

Mouse Model of Surgically-induced Endometriosis by Auto-transplantation of Uterine Tissue

Endometriosis is a chronic, painful disease whose etiology remains unknown. Furthermore, treatment of endometriosis can require laparoscopic removal of lesions, and/or chronic pharmaceutical management of pain and infertility symptoms. The cost associated with endometriosis has been estimated at 22 billion dollars per year in the United States1. To further our understanding of mechanisms underlying this enigmatic disease, animal models have been employed. Primates spontaneously develop endometriosis and therefore primate models most closely resemble the disease in women. Rodent models, however, are more cost effective and readily available2. The model that we describe here involves an autologous transfer of uterine tissue to the intestinal mesentery (Figure 1) and was first developed in the rat3 and later transferred to the mouse4. The goal of the autologous rodent model of surgically-induced endometriosis is to mimic the disease in women. We and others have previously shown that the altered gene expression pattern observed in endometriotic lesions from mice or rats mirrors that observed in women with the disease5,6. One advantage of performing the surgery in the mouse is that the abundance of transgenic mouse strains available can aid researchers in determining the role of specific components important in the establishment and growth of endometriosis. An alternative model in which excised human endometrial fragments are introduced to the peritoneum of immunocompromised mice is also widely used but is limited by the lack of a normal immune system which is thought to be important in endometriosis2,7. Importantly, the mouse model of surgically induced endometriosis is a versatile model that has been used to study how the immune system8, hormones9,10 and environmental factors11,12 affect endometriosis as well as the effects of endometriosis on fertility13 and pain14.

A description of the surgical induction of endometriosis in mice and rats by auto-transplantation of uterine tissue to the arterial cascade of the intestinal mesentery.

自动子宫组织移植手术诱发子宫内膜异位症小鼠模型

Endometriosis is a chronic, painful disease whose etiology remains unknown. Furthermore, treatment of endometriosis can require laparoscopic removal of ...

科研

医学

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

伤口愈合的小鼠模型

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

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 ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

蛛网膜下腔出血的小鼠模型

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

A Preclinical Murine Model of Hepatic Metastases

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, murine pancreatic tumor model of hepatic metastases via a hemispleen injection of syngeneic murine pancreatic tumor cells is described. This model mimics many of the clinical conditions in patients with metastatic disease to the liver. Mice consistently develop metastases in the liver allowing for investigation of the metastatic process, experimental therapy testing, and tumor immunology research.

A preclinical, murine model of hepatic metastases performed via a hemispleen injection technique.

肝转移的临床前小鼠模型

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

A Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

一个简单的临界尺寸股骨缺损模型小鼠

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of cancer metastasis are still unclear. Animal models are essential for elucidating the mechanisms and for evaluating novel strategies for the treatment of metastatic cancers. Here, an in-depth description of a &#8220;patient-like&#8221; orthotopic syngeneic mouse model for exploring the mechanisms of metastasis of solid organ tumors is provided. The survival surgical implantation of BNL 1ME A.7R.1 mouse hepatocellular carcinoma cells directly into the liver (the organ of origin) of the inbred wild-type immune competent laboratory mouse strain, BALB/c is described. The success and reproducibility of this methodology recommends it for widespread use in elucidating the biological mechanisms of solid organ cancer metastasis.

The metastatic spread of cancer is the major cause of cancer-related deaths. We provide an in-depth description of our survival surgery methodology for establishing a &#8220;patient-like&#8221; orthotopic syngeneic mouse model system for studying the mechanisms of metastasis in solid organ tumors.

"以病人为像"肝癌原位同源小鼠模型的癌转移

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of ...

Injection of Syngeneic Murine Melanoma Cells to Determine Their Metastatic Potential in the Lungs

Approximately 90% of human cancer deaths are linked to metastasis. Despite the prevalence and relative harm of metastasis, therapeutics for treatment or prevention are lacking. We report a method for the establishment of pulmonary metastases in mice, useful for the study of this phenomenon. Tail vein injection of B57BL/6J mice with B16-BL6 is among the most used models for melanoma metastases. Some of the circulating tumor cells establish themselves in the lungs of the mouse, creating "experimental" metastatic foci. With this model it is possible to measure the relative effects of therapeutic agents on the development of cancer metastasis. The difference in enumerated lung foci between treated and untreated mice indicates the efficacy of metastases neutralization. However, prior to the investigation of a therapeutic agent, it is necessary to determine an optimal number of injected B16-BL6 cells for the quantitative analysis of metastatic foci. Injection of too many cells may result in an overabundance of metastatic foci, impairing proper quantification and overwhelming the effects of anti-cancer therapies, while injection of too few cells will hinder the comparison between treated and controls.

Metastasis plays a profound role in the virulence of cancer, accounting for an estimated 90% of deaths. We report a protocol for a metastatic melanoma model in mice that is useful for determining the efficacy of therapeutic agents against this clinical phenomenon.

同基因小鼠黑色素瘤细胞注射，以确定其在肺转移潜能

Approximately 90% of human cancer deaths are linked to metastasis.  Despite the prevalence and relative harm of metastasis, therapeutics for treatment or ...

Murine Distal Colostomy, A Novel Model of Diversion Colitis in C57BL/6 Mice

Diversion colitis (DC) is a frequent clinical condition occurring in patients with bowel segments excluded from the fecal stream as a result of a diverting enterostomy. The etiology of this disease remains ill-defined but appears to differ from that of classical inflammatory bowel diseases such as Crohn's disease and ulcerative colitis. Research aimed to decipher the pathophysiological mechanisms leading to the development of this disease has been severely hampered by the lack of an appropriate murine model. This protocol generates a murine model of DC that facilitates the study of the immune system's role and its interaction with the microbiome in the development of DC. In this model using C57BL/6 animals, distal parts of the colon are excluded from the fecal stream by creating a distal colostomy, triggering the development of mild to moderate inflammation in the excluded bowel segments and reproducing the hallmark lesions of human DC with a moderate systemic inflammatory response. In contrast to the rat model, a large number of genetically-modified murine models on the C57BL/6 background are available. The combination of these animals with our model allows the potential roles of individual cytokines, chemokines, or receptors of bioactive molecules (e.g., interleukin (IL)-17; IL-10, chemokine CXCL13, chemokine receptors CXCR5 and CCR7, and the sphingosine-1-phosphate receptor 4) to be assessed in the pathogenesis of DC. The availability of congenic mouse strains on the C57BL/6 background largely facilitates transfer experiments to establish the roles of distinct cell types involved in the etiology of DC. Finally, the model offers the opportunity to assess the influences of local interventions (e.g., modification of the local microbiome or local anti-inflammatory therapy) on mucosal immunity in affected and non-affected bowel segments and the on systemic immune homeostasis.

Murine distal colostomy provides a murine model for human diversion colitis, a predominantly lymphocytic colitis in colon segment excluded from the fecal stream.

小鼠远端结肠造口术, 一种新的 C57BL/6 老鼠转移性结肠炎模型

Diversion colitis (DC) is a frequent clinical condition occurring in patients with bowel segments excluded from the fecal stream as a result of a ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...