この作品は、欧州連合のホライゾン 2020 (グラント契約号 645710) と第 7 フレームワークから (グラント契約番号 612547 と 319010) のプログラムの資金を受信しました。著者生検の手順と支援の博士フォティーニ Sourli Chasioti を感謝したいです。著者らはまた、ビデオの準備で彼の貴重な助け氏 Yiorgos Paterakis を感謝します。

研究ヘルシンキ宣言によって設定された基準に適合し、運動科学部テッサリア大学 Βiοethics 委員会によって承認されました。
 1 です手術機器や消耗品の準備 <ol><li>4輪手術メイヨーのスタンドに消毒手術フィールドの位置。</li>。
	<li>4輪手術メイヨーのスタンドに以下の消耗品・手術用機器の位置: 
	ストレート営業はさみ (15 cm) の 1 つのペア 1 メス (11 号) 1 メス ハンドル 曲線はさみ (14 cm) の 1 つのペア 1 つモスキート鉗子 コッヘル ピンセット 1 組 1 つの外科ピンセット ポビドン ヨード (の生殖ガーゼ) One 縫合 (4.0) ストレートはさみ (11 cm) の 1 つのペア 1 つ針ホルダー (15 cm) 5 滅菌ガーゼ 1 つの接着剤滅菌ガーゼ 消毒された針を用いて 1 つの 10 mL シリンジ 2% キシロカイン (ないアドレナリン) 10 mL 1 つ消毒手術用手袋のペア</li> <li>別のベンチに以下の材料を位置: 2 つのマイクロ遠心チューブ用 10% ホルマリン 5 mL を含む 1 つの管 50-100 mL の-190 ° C で液体窒素を含んでいる 1 つの小さい容器</li></ol> 2。参加者の準備 <ol><li>参加者 15 生検の手順の前に 8 時間高速に従います</li>。
	<li>参加者を控える運動、過度のストレス、アルコール、およびアクティブおよび受動喫煙 72 h 前生検 16 , 17 を持っている</li>。
</ol> 3。皮下脂肪組織の生検を行う <ol><li>仰臥位で手術のベッドの上、参加者を配置します</li>。
	<li>ポビドン ヨードを使用して切開部を消毒します</li>。
	<li>ゆっくりと着実に 2% キシロカイン (ないアドレナリン) 10 mL を注入局所麻酔として使用する選択した領域に 18.</li>
	<li>切開部を除く参加者の体をカバーする領域上で滅菌手術フィールドを配置します</li>。
	<li>少なくとも 3-5 分は、その後、ピンセットのペアを持つ局所麻酔の領域に圧力を適用、彼の感じているその局所麻酔が達成されていることを確認する任意の痛み 19 参加者を求める</li>。
	<li>号 11 メスとメス ハンドルを付ける</li>。
	<li>へその右または長さ 2 〜 2.5 cm、左に 3-5 cm の切開を作成します</li>。
	<li>営業はさみストレート 15 cm のペアで皮下組織を削除します</li>。
	<li>脂肪組織を明らかにすると、一度はピンセットのペアと湾曲した 14 cm はさみのペアでそれをキャプチャします</li>。
	<li>カット約 1 g の脂肪組織を削除します</li>。
	<li>次の組織の除去、止血して切開に滅菌ガーゼを置きます</li>。
	<li>止血を確認すると、キャプチャ 14 cm 針で 4.0 の縫合と皮膚外科ピンセットのペアと皮下組織が表示されるまでします</li>。
	<li>皮膚や皮下に密接に並べながら皮下組織へ移動する、皮膚に縫合針を挿入します</li>。
	<li>縫合針、皮下組織、皮膚を外部から駆動するまでに連続ジグザグ法に従います。皮膚切開が閉じられるまで継続します</li>。
	<li>外部から縫合糸を結ぶ</li>。
	<li>生理食塩水で切開部をきれいにし、接着剤の滅菌ガーゼでカバーします</li>。
</ol> 4。組織の沈着 <ol><li>3枚に収集した脂肪組織を分ける: 150、350 ω、500 mg</li>。
	<li>マイクロ遠心チューブ用のサンプル組織の 150 および 350 mg の作品を配置します</li>。
	<li>-80 で冷凍庫の最終的な沈着まで-190 ° C で液体窒素で小型の容器にマイクロ遠心チューブを用ひたる 2 ° C 
	注: 事前に-190 で液体窒素中でマイクロ遠心チューブ用の浸漬中カップでの失敗を避けるためにマイクロ遠心チューブ用の小さな穴をパンクに滅菌針を使用 ° C</li>
	<li>500 mg のサンプル組織片を浸し-組織学的解析に使用する-10% ホルマリン 5 mL を含む管内 。
	注: は、手術中に出血した場合にのみモスキート鉗子を使用します。この場合において、結紮して止血が達成されるまでを閉じるモスキート鉗子で船をキャプチャします</li>。
</ol>

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著者は、彼らは競合する金銭的な利益があることを宣言します。

本研究の目的は、健康な男性の非ジアテルミー法を用いた局所麻酔下で実行、腹部皮下脂肪の生検のための手術手技の標準化をだった。ジアテルミー、オペレーティング ルームで使用されます。ただし、可能性のある副作用 (すなわち、フラッシュ火災と環境毒性)9と10の治癒の遅れが発生します。ジアテルミーのこれらの不必要な効果は、営業職員...

脂肪組織生検は、代謝と疾患に関するメカニズムの洞察力に富んだ概要を提供可能性があります分析、組織サンプルを提供しています。例えば、乳房癌腫瘍同定1、脂肪酸組成試験2、重要な代謝とホルモンの病気経路3の研究に使用されます。腹部の皮下脂肪組織生検を得るためには、研究者や医師は、手術または針ベースの手法を使用します。しかし、手術皮下脂肪の生検は、複雑な生物学的メカニズム[メッセンジャーのリボ核酸 (mRNA) 式など、組織学的解析等]のより包括的な概要を提供することが組織サンプルを提供できます4.彼らは健康な成人4で使用されている以前とベッドサイド5で安全に実行できます。
針による生検法は組織の整合性を破壊する強烈な脂肪吸引を必要とし、それは通常組織 (100-500 mg)6の少量を取得します。これら組織サンプルは、複雑な研究のために適切かもしれない。さらに、針ベースの手法と比較して、手術皮下脂肪の生検を提供できる (例えば、 B および T 細胞シグナリング); いくつかの炎症性指標のより完全な絵を描くことがあります組織サンプル細胞骨格の調節;携帯電話経路;脂質、炭水化物およびアミノ酸の代謝経路酸化的リン酸化経路4。また、針によるには組織4,報7,8 の線維化領域について少し情報を提供しながらの手術 1 つ4より血液細胞汚染のリスクが高いが含まれて.
通常、手術の脂肪組織生検には9過剰な出血を防ぐために血管を焼灼のジアテルミー治療が含まれます。ジアテルミーは、燃やすし、腫瘍や疣贅病9で感染している組織によって影響を受ける組織の破壊にも使用されます。この手法は、特に脳神経や目の手術9に該当します。
ジアテルミーの過度の使用は、壊死組織、感染の原因と治癒過程10を遅らせることの形成につながる可能性があります。確かに、次のジアテルミー9フラッシュ火災など皮膚病変組織の副作用を報告されています。さらに、参加者/患者と営業スタッフ9によって吸入されることがあります手術の環境放出のジアテルミー手術煙の毒性について懸念を提起されています。確かに、ジアテルミーの使用、喫煙 6 タバコ11によって生成された同じ手術室空気毒性を引き起こす可能性があり、少なくとも 20 分12 汚染することができます最新の手術室は通常、十分な換気システムを持っていますが、.しかし、扁桃手術患者、ジアテルミー法少ないが、有意ではない、術後の痛み13と古典的な結紮止血法より出血14は示します。全体的にみて、手術後の痛みの違い、かろうじて健常者に脂肪組織生検後の率の出血の証拠が存在します。証拠の欠如とジアテルミーの副作用を考えると、脂肪組織の生検のため非ジアテルミー法の標準化が必要です。したがって、本研究の目的は健康な男性で非ジアテルミー法を用いた局所麻酔下で腹部の皮下の脂肪組織生検のための手術手技の標準化をだった。さらに、我々 は組織コレクション mRNA の発現および蛋白質の集中を決定し、組織学的解析を実行するための手順を概説します。

<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

脂肪組織生検は、代謝と疾患に関するメカニズムの洞察力の概要を提供することが分析、組織サンプルを提供しています。腹部の皮下脂肪組織生検を得るためには、研究者や医師は、手術または針ベースの手法を使用します。しかし、手術皮下脂肪の生検は白色脂肪組織の生物学的指標の複雑さのより包括的な概観を提供することが組織サンプルを提供できます。通常、手術の脂肪組織生検には、過度の出血を防ぐために血管を焼灼ジアテルミー治療が含まれています。それにもかかわらず、ジアテルミー後フラッシュ火災、体内、皮膚病変などの副作用が報告されています。したがって、非ジアテルミー法を用いた局所麻酔下で腹部の脂肪組織を外科的生検の標準化を目的とします。非ジアテルミー腹部外科的生検法による健康な男性で 115 の皮下脂肪の生検を行った.我々 の結果は、115 事業 (2.61%) から出血過剰なポスト処理の 3 つのケースを示した。結論としては、最小限の副作用で、ベッドサイドでの健康的な男性に非ジアテルミー メソッドを使用して、標準化された腹部皮下脂肪外科的生検を安全に適用できます。

参加者の特性は、表 1に提供されます。切開部の縫合は、8-12 日以内から落ちた。我々 は健常成人男性で非ジアテルミー腹部外科的生検法 115 の皮下脂肪組織生検の合計を完了した (年齢: 35.2 ± 6.9 年; ボディマス指数: 27.3 ± 4.4; および体脂肪率: 27.0 ± 9.0%)。3 つのケースで観察された後生検出血 (有病率: 2.61%)、手術後 2 日間。過度の出血を経験した 3 人は?...

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

我々 は局所麻酔下で非ジアテルミー法を用いた腹部脂肪組織生検を標準化しました。115 事業 (2.61%) から出血過剰な術後の症例が発生しました。健康な男性に非ジアテルミー法を用いた腹部脂肪組織の外科的生検を安全に適用できることが分かった.

非ジアテルミー法による腹部皮下脂肪の生検のためのテクニック

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 ビュー.  University of Thessaly. 我々 は局所麻酔下で非ジアテルミー法を用いた腹部脂肪組織生検を標準化しました。115 事業 (2.61%) から出血過剰な術後の症例が発生しました。健康な男性に非ジアテルミー法を用いた腹部脂肪組織の外科的生検を安全に適用できることが分かった.

ビデオ: 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.

間褐色脂肪組織のチェレンコフ発光イメージング

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

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.

変更されたベルイストローム法を用いたヒト骨格筋生検手順

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.

経皮針生検によって得られた骨格筋組織から単離されたミトコンドリアの調製と呼吸計測評価

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.

全組織生検からヒト上皮EnteroidsとColonoidsの確立

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.

移植のためのカプセル化サーモ前脂肪細胞の脂肪組織デポへ

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.

浸透圧を使用して皮下アンジオテンシンII注入は、マウスで誘導大動脈瘤パンプス

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.

マウスの腹部癒着の作成

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.

外ヴァーサ栄養血管の血流をブロックすることにより腹部の大動脈瘤の齧歯動物モデルの作成

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.

マウス島の口内皮下白色脂肪組織(ISWAT)移植モデル

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.

移植質評価のための化学バイオプシーの使用

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