カーン博士は、NIH/NICHD K12HD052892、アルファ 1 の基礎、ヒルマン財団およびピッツバーグ肝臓研究センターから助成を認めています。博士 Michalopoulos は、NIH/NIDDK P01DK096990 から助成を認めています。アン ・ オアの優秀で、寛大な技術的な支援お願い申し上げます。マイクロサージャリー移植または齧歯動物ラボへのアクセスを提供するためも博士デビッド A. ゲラーに感謝しています。

動物介護制度とピッツバーグ大学の利用委員会の倫理的なガイドラインに従ってすべてのプロシージャを行った。
1. 基本的なテクニックと一般的な手順
<ol><li>オートクレーブ使用する前にすべての顕微鏡装置。</li>
	<li>麻酔によりマウスを殺すことを避けるため、慎重にその呼吸パターンを確認します。</li>
	<li>臓器や器官を損傷し、出血を引き起こしているを避けるために可能な限り少し腸をタップします。湿式の綿棒やガーゼで出血点を押してください。</li>
	<li>臓器の穏やかな操作にウェット綿棒を使用します。2 つのサイズでウェット、非織ガーゼを使用、3.5 × 3.5 cm と 6.0 × 6.0 cm。 は全肝を撤回する小さいものを使用します。大きい 1 つを使用して、小規模および大規模な腸をラップします。</li>
</ol>2. 術前準備
<ol><li>男性の c57/b6 マウス部分胆管結紮用 (体重が約 20 35 g) 時代の 8 〜 12 週間を使用します。</li>
	<li>麻酔導入までマウス水や食料への無料アクセスを許可します。</li>
</ol>3. 部分胆管結紮操作
<ol><li>1.5 から 2.0 パーセントのイソフルランと 1 L/分の酸素を混ぜた誘導の商工会議所に動物を置くことによってマウスを麻酔します。呼吸を観察することによって動物を注意深く監視します。動物の呼吸速度は 1 秒あたり 1 つの息をマウスがよく麻酔です。以来、前肢逃避反射はより急速に消え、逃避反射の完全な不在を確認する同様に後肢の足をピンチします。目は、角膜潰瘍を防ぐために眼科用潤滑剤と潤滑する必要があります。メモその鎮痛剤の投与は痛みの受容体の刺激を防ぐために手術前に与えることができます。</li>
	<li>シェイブ バリカン、腹壁、手術用顕微鏡下で手術のテーブルの上にマウスを置きます。テーブルにすべての手足をテープで固定します。イソフルラン吸入を全身麻酔下でマウスを維持するために使用します。<ol>
<li>イソフルラン気化器を調整し、誘導 2.0 3.0% イソフルランを使用します。</li>
	</ol>
</li>
	<li>腹部壁の消毒滅菌綿棒を使用して、最初 0.3 0.5 mL ポビドン ヨードの局所的に適用は 70% のエタノールが続きます。動物は、滅菌ドレープで覆われるべきであります。</li>
	<li>はさみとアドソン鑷子を使用して恥骨に、剣から正中腹部切開を確認します。0.8 1.0 にイソフルランを減らす腹部切開をしたら、メンテナンスのため %。</li>
	<li>マイクロ リトラクターを使用して腹腔内を公開します。モスキート鉗子で、剣を押し、マウスの頭に向かって撤回します。ソフト粘土を使用して場所のモスキート鉗子を修正します。</li>
	<li>温めた (37 ° C) の 1-2 の mL を植え付ける滅菌リン酸緩衝生理食塩水 (PBS) 腹部臓器が負傷を避けるために腹腔内に。濡れた綿棒で小腸を押し軽くマウスの頭に向かってそれらを撤回します。小腸、小腸 (図 1 a) を負傷を避けるために後腹膜間靭帯をカットします。</li>
	<li>ウェット、不織布-ガーゼ スポンジでの小規模および大規模な腸をラップします。腹腔内の尾側に配置します。滅菌ガーゼに包まれた腸は、汚染を避けるため無菌ドレープ フィールドを配置できます。</li>
	<li>優しく肝門部、肝十二指腸靭帯 (図 1 b) を公開する小型湿式不織布-ガーゼ スポンジを使用して、剣に向かって全体の肝臓を撤回します。</li>
	<li>肝門部の解剖を確認を確認します。総胆管は門脈の腹側にあります。肝門部の主な胆道合流地点に流出する右および左の肝の胆管の位置を確認します。</li>
	<li>左葉を排水左胆管を識別し、縫合針 (図 2 a) の鈍側を使用して 10-0 ナイロン縫合糸とそれを取り囲みます。避けるために門脈、肝動脈、肝表面を負傷します。</li>
	<li>10-0 ナイロン左胆管を結紮します。(図 2 b)。</li>
	<li>湿式の綿棒を使用して、操作の前と同じ位置で小腸・大腸を配置します。</li>
	<li>腸管の捻転と腹腔内出血がないことを確認し、筋肉と 4-0 vicryl の二層で皮膚を閉じます。皮膚縫合用編組縫合 (Vicryl) が使用されたが、感染の可能性を減らすためにモノフィラメントを優先します。 注: は、恥骨に剣から継続的な縫合糸で筋膜を閉じて剣に恥骨から継続的な縫合で皮膚を閉じます。</li>
	<li>切開部を閉じる後麻酔を停止します。</li>
</ol>4. 術後の治療とフォロー アップ
<ol><li>ちょうど手術後がマウスの背側の皮をつまむし、注入 Buprenex (0.1 mg/kg) とセファゾリン (100 mg/kg 皮下、それぞれ Buprenex 注入による 12 時間間隔 3 日、セファゾリンのすべての 24 h 3 日間続いた11,12 ,13。メモその鎮痛剤の投与は痛みの受容体の刺激を防ぐために手術前に与えることができます。セファゾリンと他の周術期の抗生物質だけ移植実験で使われたもののような免疫抑制動物に必要なです。 注: 場合手術を正常に実行すると、マウス通常回復 30 分以内、マウスの活動は術前の状態とほぼ同じになります。</li>
	<li>毎日苦痛の兆候のマウスを確認してください。<ol>
<li>PBDL 後肝臓と (すなわち7、14、21 日) 定義された時点で他の組織サンプルを調達する全身麻酔下頚部転位によってマウスを犠牲に。</li>
	</ol>
</li>
</ol>

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著者の開示があります。

CBDL は閉塞性胆汁うっ滞のため最も一般的な実験的手法が、齧歯動物の複数の課題が発生することができます。術後日のエンドポイントを達成するために必要な数、に応じて、動物は重要な罹患率と時間が進むにつれて死亡率を体験できます。肝臓病の異常肝実質とベースラインで関数を示すいくつかのトランスジェニック マウス モデルを操作するとき、組織の損傷および胆管壊死の程度が...

急性肝障害・肝部分切除や胆管結紮 (cBDL) などの修復手術のモデルは、肝再生と病理学1,2を研究する齧歯動物で何十年も使用されています。CBDL で (これは肝臓で生成されるすべての胆汁を排出) 胆管結紮は、肝硬変3に最終的な進行すると術後最初の 2 〜 3 週間で閉塞性胆汁うっ滞、炎症および線維化の結果します。胆管の増殖、分化転換、および居住者の肝前駆細胞の誘導 cBDL4、5の機能があります。CBDL は、再現可能な成果野生型マウスと正常な肝臓6,7,8でこれらの系統の特徴である閉塞性 cholangiopathy の特定のメカニズムを解明し、プロシージャは、肝疾患、高い罹患率と死亡率は時間の経過とともに胆道閉塞のため予想よりもより多くの動物を必要とするいくつかの遺伝子導入マウスモデルで技術的に厳しいことができます。この手法も長時間コース実験 cBDL のストレスをそれ以外の場合は許せないかもしれないトランスジェニック マウスの基になると肝疾患の勉強のために有益です。
pBDL は、いくつかのこれらの課題を克服するために合理的な選択肢を提供できます。野生型マウスの初期 pBDL 研究は、炎症9の演技のローカルおよび全身の仲介人を区別するために目指しています。PBDL、総胆管の上左右の肝胆汁管の主な胆道合流は慎重に、門で可視化し、ローカライズされた閉塞性胆汁うっ滞を生成するのみ、左肝内胆管を結紮します。pBDL では、cBDL にいくつかの利点を提供しています。PBDL、unligated の右葉は栄養状態や要因を循環など、同じ全身作用を受ける結紮の左葉の同じ牌内部統制として機能します。全身性炎症性メディエーターを調べたところ、大沢ら以下の線維化および unligated の葉の壊死が観察が、炎症細胞と変形の成長因子 β 式結紮葉9に増加します。
最近では、pBDL が保持されて胆汁10の肝実質の効果を研究する再定義されました。(Unligated「シャム」) 内部統制群と実験群の同じ動物を使用して、pBDL は効果的により費用対効果は実験ごとに必要な動物の数を半減します。マウスは、コース実験理想的な時間 (≥2 週) 結紮葉で達成することができますので、はるかに良い、pBDL の影響を容認します。最も重要なは、pBDL は閉塞性胆汁うっ滞の直接的な肝内効果を区別することができます、unligated コントロール葉に比べて、結紮葉の胆汁構成成分を保持します。全体的に、pBDL は肝臓でローカライズされた胆汁うっ滞の葉固有の効果を研究する魅力的な方法です。

<table><tbody><tr><td>Microscope-Leica Wild M650, 6&amp;ndash;40&amp;times; magnification</td><td>Leica Microsystems</td><td>n/a</td><td></td></tr><tr><td>Tec 3 isoflurane funnel fill vaporizer</td><td>General Anesthetic Services</td><td>n/a</td><td></td></tr><tr><td>Stevens Tenotomy Scissors</td><td>Accurate Surgical &amp;amp; Scientific Instruments Corporation</td><td>ASSI.9193</td><td></td></tr><tr><td>Adson Forceps</td><td>Fine science tools</td><td>11006-12</td><td></td></tr><tr><td>Mosquito classic delicate hemostatic forceps</td><td>Codman</td><td>30-4472</td><td></td></tr><tr><td>Micro-retractor</td><td>Murdock; Roboz Surgical Instrument</td><td>RS-6550</td><td></td></tr><tr><td>Nonwoven gauze sponges</td><td>Fisherband</td><td>870-PC-DBL</td><td></td></tr><tr><td>Puritan 6&amp;rdquo; Small Cotton Swab w/Wooden Handle</td><td>Puritan Medical Products Company</td><td>868-WCS</td><td></td></tr><tr><td>Dumont SS Forceps - Standard Tips/Straight/Inox/13.5cm</td><td>Fine science tools</td><td>11203-23</td><td></td></tr><tr><td>Dumont SS Forceps - Standard Tips/Angled 45&amp;deg;/Inox/13.5cm</td><td>Fine science tools</td><td>11203-25</td><td></td></tr><tr><td>Vannas Spring Scissors - 3mm Cutting Edge</td><td>Fine science tools</td><td>15000-00</td><td></td></tr><tr><td>Microneedle holder</td><td>Aesculap Surgical Instruments</td><td>FD231R</td><td></td></tr><tr><td>Micro AROSuture, sterile 10-0 nylon suture, 70 &amp;micro;m, TAP points</td><td>AROSurgical Instruments Corporation</td><td>T4A10N07</td><td></td></tr><tr><td>4-0 Vicryl</td><td>Ethicon</td><td>J662H</td><td></td></tr><tr><td>5-ml Syringe</td><td>BD</td><td>309646</td><td></td></tr><tr><td>Falcon tissue culture dish, 60 &amp;times; 15 mm</td><td>Corning</td><td>353002</td><td></td></tr><tr><td>Isoflurane, United States Pharmacopeia (USP) liquid for inhalation, 250 ml</td><td>Piramal Healthcare</td><td>NDC 66794-017-25</td><td></td></tr><tr><td>Buprenex injectable (buprenorphine hydrochloride)</td><td>Reckitt Benckiser Pharmaceuticals</td><td>NDC 12496-0757-1</td><td></td></tr><tr><td>Cefazolin for injection , USP</td><td>APP Pharmaceuticals</td><td>NDC 63323-238-61</td><td></td></tr><tr><td>PVP scrub solution (povidone&amp;ndash;iodine 7.5%)</td><td>Medline</td><td>NDC 12496-0757-1</td><td></td></tr><tr><td>70% Ethanol</td><td>Decon Labs</td><td>2716</td><td></td></tr><tr><td>Phosphate Buffered Saline Without Calcium or Magnesium</td><td>LONZA</td><td>17-516F</td><td></td></tr><tr><td>Sirius Red</td><td>Sigma</td><td>365548</td><td></td></tr><tr><td>Hematolxylin</td><td>Fisher (Richard-Allan Scientific)</td><td>22-050-111 (7211)</td><td></td></tr><tr><td>Eosin</td><td>Anatech (Fisher)</td><td>832 (NC9686037)</td><td></td></tr><tr><td>Formalin</td><td>Fisher</td><td>SF100-20</td><td></td></tr></tbody></table>

partial bile duct ligation in the mouse: a controlled model of localized obstructive cholestasis

総胆管の完全な胆管結紮 (cBDL) は、齧歯動物、閉塞性胆汁うっ滞、肝内胆管の増殖を研究するための確立された手術方法です。ただし、長期的な実験は増加罹患率と死亡率につながることができます。選択マウス系統基礎疾患、単一動物の損失および費用を減らすことができます肝葉の結紮とも有意義な比較を可能です。ここで、説明部分胆管結紮 (pBDL) にのみ左肝内胆管を結紮は、マウスの左葉ですがない、残りの葉の胆管閉塞の原因します。同じ動物で同じ条件に服従するときに unligated の葉は、結紮葉に内部統制として役割を果たすため注意して顕微鏡手術と pBDL 実験が費用対効果をすることができます。CBDL とは異なり、別の偽手術群は必要ではありません。pBDL、胆汁うっ滞及び他の保有胆汁成分の全身影響とローカライズされた直接の比較に非常に役立ちます。pBDL は薬や細胞遊走に関連するメカニズムを調査する手法として用途に使用することができます。

この典型的な実験のため pBDL は、α 1 アンチトリプシン (A1AT) 欠乏は、人間の肝臓病を要約する帳票のピッツ ・ トランスジェニック マウスで行われました。PiZ マウス染色遺伝子の DNA の突然変異人間 A1AT 遺伝子、プロモーター、イントロンと上流と下流地域14の側面の ~ 2 kilobases のコーディング領域を含む複数のゲノム断片から成る <sup class...

Watch this Scientific Journal Video about Partial Bile Duct Ligation in the Mouse: A Controlled Model of Localized Obstructive Cholestasis at JoVE.com

Partial Bile Duct Ligation in the Mouse: A Controlled Model of Localized Obstructive Cholestasis

肝臓の傷害および齧歯動物の再生の手術モデルとして部分胆管結紮を紹介します。

マウスの部分胆管結紮: ローカライズされた閉塞性胆汁うっ滞の制御モデル

In rodents, complete bile duct ligation (cBDL) of the common bile duct is an established surgical technique for studying obstructive cholestasis and bile ...

partial-bile-duct-ligation-mouse-controlled-model-localized

Research

JoVE Journal

Biology

15.4K ビュー.  Jichi Medical University. 肝臓の傷害および齧歯動物の再生の手術モデルとして部分胆管結紮を紹介します。

ビデオ: Partial Bile Duct Ligation in the Mouse: A Controlled Model of Localized Obstructive Cholestasis

Creation of Reversible Cholestatic Rat Model

Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Cholestasis can cause various physiological changes and affect the nutritional status and surgical outcomes. Study of the pathophysiological changes occurring in the liver and other organs is of importance. Various studies have been done in cholestatic rat models.
We used a reversible cholestatic rat model in our recent study looking at the role of methylprednisolone in the ischemia reperfusion injury. Various techniques for creation of a reversible cholestatic model have been described. Creation of a reversible cholestatic rat model can be challenging in view of the smaller size and unique hepatopancreatobiliary anatomy in rats. This video article demonstrates the creation of a reversible cholestatic model.
This model can be used in various studies, such as looking at the changes in nutritional, physiological, pathological, histological and immunological changes in the gastrointestinal tract. This model can also be used to see the effects of cholestasis and various therapeutic interventions on major hepatic surgeries.

Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Creation of a reversible cholestatic rat model can be challenging in view of the smaller size and unique hepatopancreatobiliary anatomy in rats. This video article demonstrates the creation of a reversible cholestatic model.

リバーシブル胆汁うっ滞ラットモデルの作成

Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Cholestasis can cause various physiological changes and ...

医学

Determining Bile Duct Density in the Mouse Liver

Mouse is broadly used as a model organism to study biliary diseases. To evaluate the development and function of the biliary system, various techniques are used, including serum chemistry, histological analysis, and immunostaining for specific markers. Although these techniques can provide important information about the biliary system, they often do not present a full picture of bile duct (BD) developmental defects across the whole liver. This is in part due to the robust ability of the mouse liver to drain the bile even in animals with significant impairment in biliary development. Here we present a simple method to calculate the average number of BDs associated with each portal vein (PV) in sections covering all lobes of mutant/transgenic mice. In this method, livers are mounted and sectioned in a stereotypic manner to facilitate comparison among various genotypes and experimental conditions. BDs are identified via light microscopy of cytokeratin-stained cholangiocytes, and then counted and divided by the total number of PVs present in liver section. As an example, we show how this method can clearly distinguish between wild-type mice and a mouse model of Alagille syndrome. The method presented here cannot substitute for techniques that visualize the three-dimensional structure of the biliary tree. However, it offers an easy and direct way to quantitatively assess BD development and the degree of ductular reaction formation in mice.

We present a rather simple and sensitive method for accurate quantification of bile duct density in the mouse liver. This method can aid in determining the effects of genetic and environmental modifiers and the effectiveness of potential therapies in mouse models of biliary diseases.

マウス肝臓における胆管密度の決定

Mouse is broadly used as a model organism to study biliary diseases. To evaluate the development and function of the biliary system, various techniques ...

発生生物学

Bile Duct Ligation in Mice: Induction of Inflammatory Liver Injury and Fibrosis by Obstructive Cholestasis

In most vertebrates, the liver produces bile that is necessary to emulsify absorbed fats and enable the digestion of lipids in the small intestine as well as to excrete bilirubin and other metabolic products. In the liver, the experimental obstruction of the extrahepatic biliary system initiates a complex cascade of pathological events that leads to cholestasis and inflammation resulting in a strong fibrotic reaction originating from the periportal fields. Therefore, surgical ligation of the common bile duct has become the most commonly used model to induce obstructive cholestatic injury in rodents and to study the molecular and cellular events that underlie these pathophysiological mechanisms induced by inappropriate bile flow. In recent years, different surgical techniques have been described that either allow reconnection or reanastomosis after bile duct ligation (BDL), e.g., partial BDL, or other microsurgical methods for specific research questions. However, the most frequently used model is the complete obstruction of the common bile duct that induces a strong fibrotic response after 21 to 28 days. The mortality rate can be high due to infectious complications or technical inaccuracies. Here we provide a detailed surgical procedure for the BDL model in mice that induce a highly reproducible fibrotic response in accordance to the 3R rule for animal welfare postulated by Russel and Burch in 1959.

Disruption of bile flow results in severe inflammatory cholestatic liver injury with a characteristic time-dependent sequence of morphological alterations. Here we present a protocol for the surgical ligation of the common bile duct in mice that allows to induce a strong fibrotic response after 21 to 28 days.

マウスにおける胆管結紮：閉塞性胆汁うっ滞による炎症性肝障害と線維症の誘発

In most vertebrates, the liver produces bile that is necessary to emulsify absorbed fats and enable the digestion of lipids in the small intestine as well ...

A Silver Nanoparticle Method for Ameliorating Biliary Atresia Syndrome in Mice

Biliary atresia (BA) is a severe type of cholangitis with high mortality in children of which the etiology is still not fully understood. Viral infections may be one possible cause. The typical animal model used for studying BA is established by inoculating a neonatal mouse with a rhesus rotavirus. Silver nanoparticles have been shown to exert antibacterial and antiviral effects; their function in the BA mouse model is evaluated in this study. Currently, in BA animal experiments, the methods used to improve the symptoms of BA mice are generally symptomatic treatments given via food or other drugs. The aim of this study is to demonstrate a new method for ameliorating BA syndrome in mice by the intraperitoneal injection of silver nanoparticles and to provide detailed methods for preparing the silver nanoparticle gel formulation. This method is simple and widely applicable and can be used to research the mechanism of BA, as well as in clinical treatments. Based on the BA mouse model, when the mice exhibit jaundice, the prepared silver nanoparticle gel is injected intraperitoneally to the surface of the lower liver. The survival status is observed, and biochemical indicators and liver histopathology are examined. This method allows a more intuitive understanding of both the establishment of the BA model and novel BA treatments.

This article describes in detail a method based on silver nanoparticles for ameliorating biliary atresia syndrome in an experimental biliary atresia mouse model. A solid understanding of the reagent preparation process and the neonatal mouse injection technique will help familiarize researchers with the method used in neonatal mouse model studies.

マウスにおける胆道閉鎖症症候群を改善するための銀ナノ粒子法

Biliary atresia (BA) is a severe type of cholangitis with high mortality in children of which the etiology is still not fully understood. Viral infections ...

免疫・感染学

Ileectomy-induced Bile Overaccumulation in Mouse Intestine

Intestinal resection is a common therapeutic approach for human diseases such as obesity, inflammatory bowel disease, Crohn's disease, and colon cancer that often results in severe short bowel syndrome-like adverse effects including bile acid diarrhea, dehydration, electrolyte disturbances, and nutrient malabsorption. Here we introduce a murine ileal resection model, termed ileectomy, to evaluate tissue communication and the maintenance of systemic homeostasis. After ileal resection, circulating blood is permanently devoid of the ileum-specific endocrine hormone fibroblast growth factor 15 (FGF15), which releases its endocrinal inhibition of bile acid synthesis in the liver. In combination with the increased production and abolished reabsorption of bile acids after removing the ileum, mice that underwent surgery suffer from bile salt overaccumulation in the intestine and associated diarrhea, morbidity, and mortality. Novel usage of the surgery model introduced in this study may provide mechanistic and functional insights into ileal control of systemic metabolic regulation in physiology and disease.

Small intestine-dependent bile acid reabsorption and feedback inhibition of hepatic bile acid synthesis is important for systemic homeostasis and health. In this study, we describe a mouse model for ileal resection to evaluate ileectomy-induced bile malabsorption, overaccumulation, and toxicity in mouse intestine.

マウス腸管で胆汁の回腸誘起蓄積

Intestinal resection is a common therapeutic approach for human diseases such as obesity, inflammatory bowel disease, Crohn's disease, and colon cancer ...

A Mouse Model of Chronic Liver Fibrosis for the Study of Biliary Atresia

Biliary atresia (BA) is a fatal disease involving obstructive jaundice, and it is the most common indication for liver transplantation in children. Due to the complex etiology and unknown pathogenesis, there are still no effective drug treatments. At present, the classic BA mouse model induced by rhesus rotavirus (RRV) is the most commonly used model for studying the pathogenesis of BA. This model is characterized by growth retardation, jaundice of the skin and mucosa, clay stools, and dark yellow urine. The histopathology shows severe liver inflammation and obstruction of the intrahepatic and extrahepatic bile ducts, which are similar to the symptoms of human BA. However, the livers of end-stage mice in this model lack fibrosis and cannot fully simulate the characteristics of liver fibrosis in clinical BA. The presented study developed a novel BA mouse model of chronic liver fibrosis by injecting 5-10 &#181;g of anti-Ly6G antibody four times, with gaps of 2 days after each injection. The results showed that some of the mice successfully formed chronic BA with typical fibrosis after the time lapse, meaning these mice represent a suitable animal model for the virus-induced liver fibrosis mechanistic study of BA and a platform for developing future BA treatments.

We established a mouse model of chronic liver fibrosis, which provides a suitable animal model for the virus-induced liver fibrosis mechanistic study of biliary atresia (BA) and a platform for future BA treatments.

胆道閉鎖症研究のための慢性肝線維症のマウスモデル

Biliary atresia (BA) is a fatal disease involving obstructive jaundice, and it is the most common indication for liver transplantation in children. Due to ...

A Mouse Model for Chronic Pancreatitis via Bile Duct TNBS Infusion

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. Several rodent models have been developed to study CP, of which the bile duct 2,4,6 -trinitrobenzene sulfonic acid (TNBS) infusion model replicates the features of neuropathic pain seen in CP. However, bile duct drug infusion in mice is technically challenging. This protocol demonstrates the procedure of bile duct TNBS infusion for generation of a CP mouse model. TNBS was infused into the pancreas through the ampulla of Vater in the duodenum. This protocol optimized drug volume, surgical techniques, and drug handling during the procedure. TNBS-treated mice showed features of CP as reflected by bodyweight and pancreas weight reductions, changes in pain-associated behaviors, and abnormal pancreatic morphology. With these improvements, mortality associated with TNBS injection was minimal. This procedure is not only critical in generating pancreatic disease models but is also useful in local pancreatic drug delivery.

Chronic pancreatitis (CP) is a disease characterized by inflammation and fibrosis of the pancreas, often associated with intractable abdominal pain. This article focuses on refining the technique to generate a mouse model of CP via bile duct infusion with 2,4,6 -trinitrobenzene sulfonic acid (TNBS).

胆管TNBS輸液による慢性膵炎のマウスモデル

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. ...

Extrahepatic Bile Duct and Gall Bladder Dissection in Nine-Day-Old Mouse Neonates

The dissection of murine neonatal bile ducts has been described as difficult. The main aim of the described standard operating procedure is the isolation of the extrahepatic bile duct (EBD) in mouse neonates without damaging the bile duct during preparation. Because of its exceptionally close preparation compared to the cholangiocytes cell line and harvesting of the entire extrahepatic bile duct system (EBDS), the described approach is extremely useful in researching animal models of newborn bile duct disorders, such as biliary atresia. After euthanasia, the peritoneal cavity was accessed, and the bile duct system, duodenum, and liver were extracted with the unique En-bloc-Resection (EbR). The extracted sample is placed on a foam mat, and the EBD is dissected from contaminating cells atraumatically without necessary touch. The dissection of the entire EBDS is a significant advantage of this method. Caution must be taken due to the small size and amount of bile duct tissue. Using the described technique, there is no damage to the cholangiocytes. Further, the purity of the technique is reproducible (n = 10). Therefore, optimally comparable samples can be harvested. Furthermore, no bile duct tissue is harmed, because any contact with the bile duct system can be avoided during preparation, leaving the bile fluid inside the gall bladder. Most importantly, while performing the final gall bladder and bile duct dissection, atraumatic microinstruments were used only slightly lateral of the bile duct without squeezing it. This is the key to a clean and intact sample, and essential for further histological investigation or the isolation of cholangiocytes. To summarize, the described innovative dissection technique enables especially inexperienced operators with the necessary equipment to isolate the EBDS as cleanly as possible.

For the observation of murine neonatal bile duct disorders, an intact bile duct and efficient preparation are required. Therefore, a new approach for isolating the entire extrahepatic bile duct system in murine neonates was successfully developed while maintaining the integrity of the bile duct.

9日齢マウス新生児における肝外胆管および胆嚢郭清

The dissection of murine neonatal bile ducts has been described as difficult. The main aim of the described standard operating procedure is the isolation ...

<meta charset="utf8"></head><body>病期的肝切除術のための動物モデルにおけるALLPSの肝臓再生の研究

A Mouse Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy Procedure Aided by Microscopy

Hepatectomy is widely regarded as the primary treatment for hepatic malignancies; yet, postoperative liver failure remains a major cause of perioperative mortality, severely impacting patient outcomes. In a robust hepatic environment, the future liver remnant (FLR) must exceed 25%, and in cases of cirrhosis, this requirement increases to over 40%. The inadequacy of FLR is currently a major obstacle in the progression of hepatic surgery.
Traditional methods to enhance FLR hypertrophy mainly focus on portal vein embolization (PVE), but its effectiveness is considerably limited. In recent years, there have been numerous reports on a novel biphasic hepatectomy method involving hepatic partitioning and portal vein ligation, known as associating liver partition and portal vein ligation for staged hepatectomy (ALPPS). ALPPS surpasses PVE in efficiently and considerably inducing FLR hypertrophy. However, the detailed mechanisms driving ALPPS-facilitated hepatic regeneration are not fully understood. Thus, replicating ALPPS in animal models is crucial to thoroughly investigate the molecular mechanisms of hepatic regeneration, offering valuable theoretical and practical insights.

<meta charset="utf8"></head><body>著者スポットライト:ALPPSマウスモデルによる肝臓再生研究の進歩

Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy (ALPPS) has been shown to result in remarkable liver regeneration after several weeks post first-stage surgery. Animal models that have undergone ALPPS have been developed to explore potential liver regeneration ability and achieve favorable clinical outcomes.

顕微鏡法による段階的肝切除術のための関連肝パーティションおよび門脈結紮のマウスモデル

Hepatectomy is widely regarded as the primary treatment for hepatic malignancies; yet, postoperative liver failure remains a major cause of perioperative ...

マウスの部分胆管結紮: ローカライズされた閉塞性胆汁うっ滞の制御モデル

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リバーシブル胆汁うっ滞ラットモデルの作成

マウス肝臓における胆管密度の決定

マウスにおける胆管結紮：閉塞性胆汁うっ滞による炎症性肝障害と線維症の誘発

マウスにおける胆道閉鎖症症候群を改善するための銀ナノ粒子法

マウス腸管で胆汁の回腸誘起蓄積

胆道閉鎖症研究のための慢性肝線維症のマウスモデル

胆管TNBS輸液による慢性膵炎のマウスモデル

9日齢マウス新生児における肝外胆管および胆嚢郭清

顕微鏡法による段階的肝切除術のための関連肝パーティションおよび門脈結紮のマウスモデル

顕微鏡法による段階的肝切除術のための関連肝パーティションおよび門脈結紮のマウスモデル