这项工作得到了欧洲联盟2020期 (赠款协议 645710) 和第七框架 (赠款协议612547和 319010) 方案的资助。作者希望感谢 Dr. Fotini Sourli-Chasioti 对活检程序的帮助。作者还感谢 Mr. Yiorgos Paterakis 在视频制作方面的宝贵帮助。

该研究符合《赫尔辛基宣言》规定的标准, 并获得萨利大学、运动科学系、#914; 我和 #959; 道德委员会. 
 1. 手术设备和消耗品的准备 <ol> <li> 将一个消毒的外科手术场放置在一个4轮手术的梅奥站上. </li>
	<li> 将下列耗材和手术设备放置在4轮手术的梅奥站上: 
	一双直操作剪刀 (15 厘米) 一手术刀 (No. 11) 一手术刀柄 一双弯曲剪刀 (14 厘米) 一只蚊子钳 一对科克镊子 一个外科镊子 聚维酮碘 (在无菌纱布) 一条缝合线 (4.0) 一双直剪刀 (11 厘米) 一针架 (15 厘米) 五无菌纱布 一胶粘剂无菌纱布 一10毫升注射器与消毒针 10mL 2% 因 (没有肾上腺素) 一消毒手术手套 </li> <li> 将以下材料放在单独的工作台上: 两个离心管 一管含有5毫升的10% 福尔马林 一个小容器, 其中含有50-100 毫升液氮在-190 和 #176;</li> </ol> 2。参加者的准备 <ol> <li> 在活检过程之前让参与者遵循8小时的快速 15 。
	<li> 让参加者在活检之前不要进行体育锻炼, 过度的压力, 酒精, 主动和被动吸烟72小时 16 , 17 . </li>
</ol> 3。执行皮下脂肪组织活检 <ol> <li> 将参与者置于仰卧位的手术床上. </li>
	<li> 使用聚维酮碘对切口区域进行消毒. </li>
	<li> 将10毫升的2% 因 (无肾上腺素) 缓慢而稳步地注入到选定区域中, 作为局部麻醉. </li>
	<li> 在该区域放置一个无菌手术场, 以覆盖参与者的身体, 不包括切口区域. </li>
	<li> 至少3-5 分钟后, 用一对镊子按压局部麻醉区域, 并询问参与者是否感到疼痛 19 以确保局部麻醉已实现. </li>
	<li> 将 No. 11 手术刀附在手术刀柄上. </li>
	<li> 创建切口 2-2. 5 厘米长度, 3-5 厘米到左边或右侧的肚脐. </li>
	<li> 用一对直15厘米的操作剪刀将皮下组织取出. </li>
	<li> 一旦脂肪组织显露, 用一对镊子和一对弯曲的14厘米剪刀捕捉它. </li>
	<li> 切割并除去大约1克脂肪组织. </li>
	<li> 组织切除后, 在切口处放置无菌纱布, 以达到止血效果. </li>
	<li> 当止血被证实时, 用14厘米针和皮肤用一双外科镊子捕捉4.0 缝合, 直到皮下组织可见. </li>
	<li> 将缝合针插入皮肤, 向皮下组织移动, 而皮肤和皮下紧密排列. </li>
	<li> 用缝合针, 沿着一个连续的曲折技术进入皮下组织, 直到从皮肤外部驱动。继续, 直到皮肤切口关闭. </li>
	<li> 外部绑住缝合线 </li>
	<li> 用生理盐水清洗切口面积, 并用无菌纱布覆盖. </li>
</ol> 4。组织的沉积 <ol> <li> 将收集的脂肪组织分离为三块: 150、350和500毫克. </li>
	<li> 将150和350毫克样品组织放置在离心管中. </li>
	<li> 将两个离心管浸入小容器中, 将液氮浸泡在-190 和 #176; c 直到在-80 和 #176 的冷冻库中最终沉积; c. 
	注: 预先使用无菌针头刺穿离心管中的一个小孔, 以避免在-190 和 #176 的液氮浸入离心管时发生杯状故障; C. </li>
	<li> 浸泡500毫克样品组织-用于组织学-一管含有5毫升10% 福尔马林. 
	注: 只有在手术过程中出血过多的情况下才使用蚊虫钳。在这种情况下, 捕获的船只与蚊子钳结扎和关闭, 直到止血达到. </li>
</ol>

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作者声明他们没有竞争的金融利益。

本研究的目的是规范一项手术技术的皮下腹部脂肪组织活检, 进行局部麻醉下进行了 non-diathermy 方法的健康男性。透在手术室里常用;但是, 可能出现的副作用 (即,闪火和环境毒性)9和愈合中的延迟10 。透的这些有害影响会给操作人员和患者/参与者带来健康风险。因此, 有必要进行皮下脂肪组织活检的手术 non-diathermy 方法。
在手术后两?...

脂肪组织活检提供组织样本, 经分析后, 可以提供有关新陈代谢和疾病的机制的有见地的概述。例如, 它们用于乳腺癌肿瘤的鉴定1, 脂肪酸组成检查的2, 以及重要的代谢和荷尔蒙疾病通路的研究3。为了获得腹部区域的皮下脂肪组织活检, 研究人员和医生使用的是外科手术或 needle-based 技术。然而, 手术皮下脂肪活检可以提供组织样本, 可能提供一个更全面的概述复杂的生物机制 [例如,信使核糖核酸 (mRNA) 的表达, 组织学分析,等]4.它们以前已用于健康成人4 , 可在床边5中安全地执行。
needle-based 活检技术需要强烈的抽脂, 可能破坏组织的完整性, 它通常检索少量的组织 (100-500 毫克)6。这些组织标本可能不适合复杂的研究研究。此外, 与 needle-based 技术相比, 手术皮下脂肪活检可以提供组织样本, 可以绘制一个更完整的图片的几个炎症指数 (例如, b-和 T 细胞信号);细胞骨架调节;细胞通路;脂质、碳水化合物和氨基酸的代谢途径;和氧化磷酸化途径4。此外, needle-based 技术包括更高的风险, 血液细胞污染比外科手术一个4, 而提供了关于组织的纤维化区域的小信息4,7,8.
通常, 手术脂肪组织活检包括透治疗烧灼血管, 以防止过多出血9。透也被用于烧伤和破坏受疾病感染的组织和疣或组织的影响9。这项技术特别适用于神经外科和眼科手术9。
过度使用透可能导致坏死组织的形成, 导致感染和延迟愈合过程10。事实上, 在透9之后, 报告的副作用如闪光火和组织中的皮肤损害。此外, 有人担心透的外科烟雾对手术环境的毒性, 这可能会被参与者/病人和操作人员吸入9。事实上, 使用透可能会导致手术室空气的毒性等于吸烟六香烟的11和, 虽然手术室通常有良好的通风系统, 污染可以持续至少20分钟12.然而, 在进行扁桃体手术的患者中, 透方法显示出较少, 但显著, 术后疼痛13和出血的14比传统的结扎止血方法。总的来说, 关于健康个体脂肪组织活检后疼痛和出血率差异的证据几乎不存在。鉴于缺乏证据和透的副作用, 有必要对脂肪组织活检的 non-diathermy 方法进行标准化。因此, 本研究的目的是规范一项手术技术的皮下腹部脂肪组织活检, 进行局部麻醉下使用 non-diathermy 法的健康男性。此外, 我们概述了组织收集的程序, 以确定 mRNA 的表达和蛋白质浓度, 并进行组织学分析。

<table border="0" cellpadding="0" cellspacing="0" width="981">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">&#160;Disinfected surgical field&#160;</td>
			<td style="width:183px;">Hartmann</td>
			<td style="width:175px;">23516-00</td>
			<td style="width:296px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stainless Steel Instrument Stands (surgical Mayo)</td>
			<td>Medical Device Depot Inc.</td>
			<td>32000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Operating Scissors straight 15 cm</td>
			<td>Karl Storz</td>
			<td>791903</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scalpel No 11&#160;</td>
			<td>Swann-Morton</td>
			<td>203</td>
			<td>Bipolar, catalog number 429011</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scarpel handle No 3</td>
			<td>Karl Storz</td>
			<td>488090</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scissors curved 14 cm</td>
			<td>Karl Storz</td>
			<td>511514, 752918</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mosquito forceps</td>
			<td>Karl Storz</td>
			<td>535012</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tweezers Kocher&#160;</td>
			<td>Karl Storz</td>
			<td>530416</td>
			<td>Bipolar, catalog number 830316</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical tweezers</td>
			<td>Karl Storz</td>
			<td>793216</td>
			<td>Bipolar, catalog number 831016</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Povidone-iodine&#160;</td>
			<td>Various</td>
			<td>Not applicable</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Suture 4.0 Vicryl</td>
			<td>Johnsons</td>
			<td>V4970H&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scissors straight 11 cm</td>
			<td>Karl Storz</td>
			<td>512511 DS</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Needle holder 15 cm</td>
			<td>Karl Storz</td>
			<td>515515</td>
			<td>Bipolar, catalog number 213015</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;Sterile gauzes</td>
			<td>Hartmann</td>
			<td>3021</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#913;dhesive sterile gauze&#160;</td>
			<td>Digas medical equipment</td>
			<td>2889</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10 mL syringe with disinfected needle</td>
			<td>Alpha medical solutions</td>
			<td>10310-55-0010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2%-xylocaine (no adrenaline)</td>
			<td>Various</td>
			<td>Not applicable</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sterile surgical gloves&#160;</td>
			<td>Sempermed</td>
			<td>10001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Eppendorf tubes</td>
			<td>Kartell</td>
			<td>16283</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10% formalin&#160;</td>
			<td>Alpha medical solutions</td>
			<td>11016-30-2909</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tubes 120 mL</td>
			<td>Digas medical equipment</td>
			<td>9025</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Liquid nitrogen at -190&#176;C</td>
			<td>Revival</td>
			<td>Not applicable</td>
			<td></td>
		</tr>
	</tbody>
</table>

a technique for subcutaneous abdominal adipose tissue biopsy via a non-diathermy method

脂肪组织活检提供组织样本, 经分析后, 可提供有关代谢和疾病机制的有见地的概述。为了获得腹部区域的皮下脂肪组织活检, 研究人员和医生使用的是外科手术或 needle-based 技术。然而, 手术皮下脂肪活检可以提供组织样本, 可以提供更全面的概述的复杂性生物指标的白色脂肪组织。通常, 手术脂肪组织活检包括透治疗烧灼血管, 以防止过度出血。然而, 在透后, 有报道称, 在组织中出现了闪光和皮肤损伤等副作用。因此, 我们的目的是规范一个外科腹部脂肪组织活检进行局部麻醉使用 non-diathermy 方法。我们进行了115皮下脂肪组织活检的健康男性使用 non-diathermy 腹部手术活检方法。我们的结果显示, 三例术后出血过多, 115 手术 (2.61%)。总之, 我们的标准化皮下腹部脂肪组织手术活检使用 non-diathermy 方法可以安全地应用于健康的人在床边, 最小的副作用。

参与者的特征在表 1中提供。切口区域的缝线在接下来的8-12 天内脱落。我们一共完成了115皮下脂肪组织活检使用 non-diathermy 腹部外科活检方法在健康成人 (年龄: 35.2 ±6.9 年; 身体质量索引: 27.3 ± 4.4; 和身体脂肪百分比: 27.0 ± 9.0%)。术后两天观察三例 (患病率: 2.61%) 后活检过度出血。三名经历过大出血的人比没有经历过任何副作用的人年长 (t(113)=-2.115, p ...

Watch this Scientific Journal Video about A Technique for Subcutaneous Abdominal Adipose Tissue Biopsy via a Non-diathermy Method at JoVE.com

A Technique for Subcutaneous Abdominal Adipose Tissue Biopsy via a Non-diathermy Method

我们用局部麻醉下的 non-diathermy 法对腹部脂肪组织进行了活检。三例术后大出血115手术 (2.61%) 发生。我们的结论是, 腹部脂肪组织手术活检使用 non-diathermy 的方法可以安全地应用于健康男性。

经非透法皮下腹脂肪组织活检技术

Adipose tissue biopsies offer tissue samples that, upon analysis, may provide insightful overviews of mechanisms relating to metabolism and disease. To ...

a-technique-for-subcutaneous-abdominal-adipose-tissue-biopsy-via-non

Research

JoVE Journal

Medicine

14.9K 次观看.  University of Thessaly. 我们用局部麻醉下的 non-diathermy 法对腹部脂肪组织进行了活检。三例术后大出血115手术 (2.61%) 发生。我们的结论是, 腹部脂肪组织手术活检使用 non-diathermy 的方法可以安全地应用于健康男性。

视频: A Technique for Subcutaneous Abdominal Adipose Tissue Biopsy via a Non-diathermy Method

Cerenkov Luminescence Imaging of Interscapular Brown Adipose Tissue

Brown adipose tissue (BAT), widely known as a &#8220;good fat&#8221; plays pivotal roles for thermogenesis in mammals. This special tissue is closely related to metabolism and energy expenditure, and its dysfunction is one important contributor for obesity and diabetes. Contrary to previous belief, recent PET/CT imaging studies indicated the BAT depots are still present in human adults. PET imaging clearly shows that BAT has considerably high uptake of 18F-FDG under certain conditions. In this video report, we demonstrate that Cerenkov luminescence imaging (CLI) with 18F-FDG can be used to optically image BAT in small animals. BAT activation is observed after intraperitoneal injection of norepinephrine (NE) and cold treatment, and depression of BAT is induced by long anesthesia. Using multiple-filter Cerenkov luminescence imaging, spectral unmixing and 3D imaging reconstruction are demonstrated. Our results suggest that CLI with 18F-FDG is a practical technique for imaging BAT in small animals, and this technique can be used as a cheap, fast, and alternative imaging tool for BAT research.

In this video report, we show the application of Cerenkov Luminescence Imaging (CLI) for interscapular brown adipose tissue in mice under activated and depressed conditions.

肩胛间褐色脂肪组织的切伦科夫发光成像

Brown adipose tissue (BAT), widely known as a &#8220;good fat&#8221; plays pivotal roles for thermogenesis in mammals. This special tissue is closely ...

科研

医学

Human Skeletal Muscle Biopsy Procedures Using the Modified Bergstr&#246;m Technique

The percutaneous biopsy technique enables researchers and clinicians to collect skeletal muscle tissue samples. The technique is safe and highly effective. This video describes the percutaneous biopsy technique using a modified Bergstr&#246;m needle to obtain skeletal muscle tissue samples from the vastus lateralis of human subjects. The Bergstr&#246;m needle consists of an outer cannula with a small opening (&#8216;window&#8217;) at the side of the tip and an inner trocar with a cutting blade at the distal end. Under local anesthesia and aseptic conditions, the needle is advanced into the skeletal muscle through an incision in the skin, subcutaneous tissue, and fascia. Next, suction is applied to the inner trocar, the outer trocar is pulled back, skeletal muscle tissue is drawn into the window of the outer cannula by the suction, and the inner trocar is rapidly closed, thus cutting or clipping the skeletal muscle tissue sample. The needle is rotated 90&#176;&#160;and another cut is made. This process may be repeated three more times. This multiple cutting technique typically produces a sample of 100-200 mg or more in healthy subjects and can be done immediately before, during, and after a bout of exercise or other intervention. Following post-biopsy dressing of the incision site, subjects typically resume their activities of daily living right away and can fully participate in vigorous physical activity within 48-72 hr. Subjects should avoid heavy resistance exercise for 48 hr to reduce the risk of herniation of the muscle through the incision in the fascia.

The purpose of this video is to present the modified Bergstr&#246;m skeletal muscle biopsy technique on human subjects.

人骨骼肌活检手术使用修改伯格斯坦技术

The percutaneous biopsy technique enables researchers and clinicians to collect skeletal muscle tissue samples. The technique is safe and highly ...

Preparation and Respirometric Assessment of Mitochondria Isolated from Skeletal Muscle Tissue Obtained by Percutaneous Needle Biopsy

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining samples of human Vastus lateralis, isolating mitochondria from minimal amounts of skeletal muscle tissue, and plate based respirometric profiling using an extracellular flux (XF) analyzer. Comparison of respirometric profiles obtained using 1.0, 2.5 and 5.0 &#956;g of mitochondria indicate that 1.0 &#956;g is sufficient to measure respiration and that 5.0 &#956;g provides most consistent results based on comparison of standard errors. Western blot analysis of isolated mitochondria for mitochondrial marker COX IV and non-mitochondrial tissue marker GAPDH indicate that there is limited non-mitochondrial contamination using this protocol. The ability to study mitochondrial respirometry in as little as 20 mg of muscle tissue allows users to utilize individual biopsies for multiple study endpoints in clinical research projects.

Methods for biopsy of Vastus lateralis, preparation of purified mitochondria, and respirometric profiling are described. The use of small muscle volume makes this technique suitable for clinical research applications.

线粒体的制备及透气性评估，从骨骼肌组织通过穿刺活检获得的分离

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining ...

Establishment of Human Epithelial Enteroids and Colonoids from Whole Tissue and Biopsy

The epithelium of the gastrointestinal tract is constantly renewed as it turns over. This process is triggered by the proliferation of intestinal stem cells (ISCs) and progeny that progressively migrate and differentiate toward the tip of the villi. These processes, essential for gastrointestinal homeostasis, have been extensively studied using multiple approaches. Ex vivo technologies, especially primary cell cultures have proven to be promising for understanding intestinal epithelial functions. A long-term primary culture system for mouse intestinal crypts has been established to generate 3-dimensional epithelial organoids. These epithelial structures contain crypt- and villus-like domains reminiscent of normal gut epithelium. Commonly, termed &#8220;enteroids&#8221; when derived from small intestine and &#8220;colonoids&#8221; when derived from colon, they are different from organoids that also contain mesenchyme tissue. Additionally, these enteroids/colonoids continuously produce all cell types found normally within the intestinal epithelium. This in vitro organ-like culture system is rapidly becoming the new gold standard for investigation of intestinal stem cell biology and epithelial cell physiology. This technology has been recently transferred to the study of human gut. The establishment of human derived epithelial enteroids and colonoids from small intestine and colon has been possible through the utilization of specific culture media that allow their growth and maintenance over time. Here, we describe a method to establish a small intestinal and colon crypt-derived system from human whole tissue or biopsies. We emphasize the culture modalities that are essential for the successful growth and maintenance of human enteroids and colonoids.

We describe a method to establish human enteroids from small intestinal crypts and colonoids from colon crypts collected from both surgical tissue and biopsies. In this methodological article, we present the culture modalities that are essential for the successful growth and maintenance of human enteroids and colonoids.

从整体组织和活检建立人类上皮Enteroids和Colonoids的

The epithelium of the gastrointestinal tract is constantly renewed as it turns over. This process is triggered by the proliferation of intestinal stem ...

Encapsulation Thermogenic Preadipocytes for Transplantation into Adipose Tissue Depots

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular weight metabolites in tissues of the treated host to achieve long-term survival. The semipermeable membrane allows engrafted encapsulated cells to avoid rejection by the immune system. The encapsulation procedure was designed to enable a controlled release of bioactive compounds, such as insulin, other hormones, and cytokines. Here we describe a method for encapsulation of catabolic cells, which consume lipids for heat production and energy dissipation (thermogenesis) in the intra-abdominal adipose tissue of obese mice. Encapsulation of thermogenic catabolic cells may be potentially applicable to the prevention and treatment of obesity and type 2 diabetes. Another potential application of catabolic cells may include detoxification from alcohols or other toxic metabolites and environmental pollutants.

Here, we present a protocol for encapsulation of catabolic cells, which consume lipids for heat production in intra-abdominal adipose tissue and increase energy dissipation in obese mice.

封装产热前脂肪细胞移植到脂肪组织仓库

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular ...

Subcutaneous Angiotensin II Infusion using Osmotic Pumps Induces Aortic Aneurysms in Mice

Osmotic pumps continuously deliver compounds at a constant rate into small animals. This article introduces a standard protocol used to induce aortic aneurysms via subcutaneous infusion of angiotensin II (AngII) from implanted osmotic pumps. This protocol includes calculation of AngII amount and dissolution, osmotic pump filling, implantation of osmotic pumps subcutaneously, observation after pump implantation, and harvest of aortas to visualize aortic aneurysms in mice. Subcutaneous infusion of AngII through osmotic pumps following this protocol is a reliable and reproducible technique to induce both abdominal and thoracic aortic aneurysms in mice. Infusion durations range from a few days to several months based on the purpose of the study. AngII 1,000 ng/kg/min is sufficient to provide maximal effects on abdominal aortic aneurysmal formation in male hypercholesterolemic mouse models such as apolipoprotein E deficient or low-density lipoprotein receptor deficient mice. Incidence of abdominal aortic aneurysms induced by AngII infusion via osmotic pumps is 5 - 10 times lower in female hypercholesterolemic mice and also lower in both genders of normocholesterolemic mice. In contrast, AngII-induced thoracic aortic aneurysms in mice are not hypercholesterolemia or gender-dependent. Importantly, multiple features of this mouse model recapitulate those of human aortic aneurysms.

Subcutaneous implantation of osmotic pumps provides a convenient approach for prolonged and consistent delivery of compounds. This approach has been used extensively to study both abdominal and thoracic aortic aneurysms in mice.

使用渗透皮下血管紧张素Ⅱ输液泵诱导主动脉瘤小鼠

Osmotic pumps continuously deliver compounds at a constant rate into small animals.  This article introduces a standard protocol used to induce aortic ...

Creation of Abdominal Adhesions in Mice

Abdominal adhesions consist of fibrotic tissue that forms in the peritoneal space in response to an inflammatory insult, typically surgery or intraabdominal infection. The precise mechanisms underlying adhesion formation are poorly understood. Many compounds and physical barriers have been tested for their ability to prevent adhesions after surgery with varying levels of success. The mouse and rat are important models for the study of abdominal adhesions. Several different techniques for the creation of adhesions in the mouse and rat exist in the literature. Here we describe a protocol utilizing abrasion of the cecum with sandpaper and sutures placed in the right abdominal sidewall. The mouse is anesthetized and the abdomen is prepped. A midline laparotomy is created and the cecum is identified. Sandpaper is used to gently abrade the surface of the cecum. Next, several figure-of-eight sutures are placed into the peritoneum of the right abdominal sidewall. The abdominal cavity is irrigated, a small amount of starch is applied, and the incision is closed. We have found that this technique produces the most consistent adhesions with the lowest mortality rate.

Abdominal adhesions that form after surgery are a major cause of pain, infertility, and hospitalization and reoperation for small bowel obstruction. Our surgical procedure for creating abdominal adhesions in mice is a reliable tool to study the mechanisms underlying the formation of adhesions.

小鼠腹腔粘连的创建

Abdominal adhesions consist of fibrotic tissue that forms in the peritoneal space in response to an inflammatory insult, typically surgery or ...

Creation of a Rodent Model of Abdominal Aortic Aneurysm by Blocking Adventitial Vasa Vasorum Perfusion

The adventitial vasa vasorum (VV) provides oxygen and nourishment to the aortic wall. Hypoxia in the aortic wall can cause enlarged abdominal aortic aneurysms (AAAs). This article introduces and describes a standard protocol used to induce AAAs through adventitial VV hypoperfusion created with a combination of polyurethane catheter insertion into the aortic lumen and suture ligation of the infrarenal abdominal aorta.
The protocol involves the use of male rats weighing 300-400 g, which are provided food and water ad libitum. After laparotomy with a ventral midline abdominal incision, exfoliation of the aorta is performed, which blocks blood flow from the perivascular tissue. Aortotomy involving a small incision adjacent to the renal artery branches is performed, and a polyurethane catheter is inserted using an 18-gauge indwelling needle. After repairing the incision, tight ligation of the aorta over the catheter blocks VV blood flow from the proximal direction through the aortic wall without disturbing the aortic blood flow. This technique can induce an AAA with progressive aortic dilatation.
The greatest benefit of this model is that VV hypoperfusion causes tissue hypoxia and the development of an infrarenal AAA, which has morphological and pathological characteristics similar to those of a human AAA.

Polyurethane catheter insertion into the aortic lumen and suture ligation of the aorta induce chronic hypoxia due to hypoperfusion of the adventitial vasa vasorum. This article describes a novel animal model of abdominal aortic aneurysm (AAA) with characteristics similar to those of AAA in humans.

阻断外瓦萨血管灌注对腹主动脉瘤鼠模型的建立

The adventitial vasa vasorum (VV) provides oxygen and nourishment to the aortic wall. Hypoxia in the aortic wall can cause enlarged abdominal aortic ...

Inguinal Subcutaneous White Adipose Tissue (ISWAT) Transplantation Model of Murine Islets

Pancreatic islet transplantation is a well-established therapeutic treatment for type 1 diabetes. The kidney capsule is the most commonly used site for islet transplantation in rodent models. However, the tight kidney capsule limits the transplantation of sufficient islets in large animals and humans. The inguinal subcutaneous white adipose tissue (ISWAT), a new subcutaneous space, was found to be a potentially valuable site for islet transplantation. This site has better blood supply than other subcutaneous spaces. Moreover, the ISWAT accommodates a larger islet mass than the kidney capsule, and transplantation into it is simple. This manuscript describes the procedure of mouse islet isolation and transplantation in the ISWAT site of syngeneic diabetic mouse recipients. Using this protocol, murine pancreatic islets were isolated by standard collagenase digestion and a basement membrane matrix hydrogel was used for fixing the purified islets in the ISWAT site. The blood glucose levels of the recipient mice were monitored for more than 100 days. Islet grafts were retrieved at day 100 after transplantation for histological analysis. The protocol for islet transplantation in the ISWAT site described in this manuscript is simple and effective.

Inguinal Subcutaneous White Adipose Tissue ISWAT Transplantation Model of Murine Islets

In this protocol, a method of murine islet isolation and transplantation into the inguinal subcutaneous white adipose tissue is described. Isolated syngeneic murine islets are transplanted into a murine recipient using a basement membrane hydrogel. The blood glucose level of the recipients is monitored, and histology analysis of the islet grafts is performed.

毛里岛皮下白脂肪组织（ISWAT）移植模型

Pancreatic islet transplantation is a well-established therapeutic treatment for type 1 diabetes. The kidney capsule is the most commonly used site for ...

Using a Chemical Biopsy for Graft Quality Assessment

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated with an increased survival rate and greater quality of patient&#8217;s life when compared to conventional dialysis. Regrettably, transplantology suffers from a lack of reliable methods for organ quality assessment. Standard diagnostic techniques are limited to macroscopic appearance inspection or invasive tissue biopsy, which do not provide comprehensive information about the graft. The proposed protocol aims to introduce solid phase microextraction (SPME) as an ideal analytical method for comprehensive metabolomics and lipidomic analysis of all low molecular compounds present in kidneys allocated for transplantation. The small size of the SPME probe enables performance of a chemical biopsy, which enables extraction of metabolites directly from the organ without any tissue collection. The minimum invasiveness of the method permits execution of multiple analyses over time: directly after organ harvesting, during its preservation, and immediately after revascularization at the recipient&#8217;s body. It is hypothesized that the combination of this novel sampling method with a high-resolution mass spectrometer will allow for discrimination of a set of characteristic compounds that could serve as biological markers of graft quality and indicators of possible development of organ dysfunction.

The protocol presents utilization of the chemical biopsy approach followed by comprehensive metabolomic and lipidomic analysis for quality assessment of kidney grafts allocated for transplantation.

使用化学活检进行移植质量评估

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated ...