説明するすべての手順を研究し、すべての動物のプロトコルにおける動物の管理と使用のための健康ガイドラインの国立研究所に準拠して実施されていますがオレゴン健康科学大学動物実験用の委員会によって承認された。 1。外科的準備<ol><li>マウスの重量を量る。説明する手順は、gの20〜25重量C57BL / 6マウスで実行されます麻酔は、イソフルラン3から4パーセントを使用してインダクションボックスに誘導し、その後、空気/酸素混合物中のイソフルラン1.5から2.5パーセントを使用して維持されます。 </li><li>目に潤滑剤を塗布し、加熱パッド上に動物の仰臥位に配置します。テープを使用して4四肢を固定化する。後肢足が中立位置で録音されることがあります、しかし、前肢は胸骨圧迫の間に完全な胸壁の遠足を可能にするために、できるだけ胸壁に近いとして保護する必要があります。 </li><li>直腸温度プローブを注油して置きます。温度は37.0を維持するために設定されている電子温度調節器（Digi - Keyはセンス、コールパーマー、バーノンヒルズIL）、℃に接続された加熱パッドとランプを使用して制御されます。それは動物内の温度勾配は、非流動状態の間に開発できる可能性があるため、それは温度を測定し、関心の器官の近くに制御することが重要です。 </li><li> 2.5センチメートル22 GAテフロンカテーテル（Insyte - W、BD、フランクリンNJ）と直角イントロ（IN Frova紹介者、クックメディカル、ブルーミントン）のカット先端を使用して気管を挿管する。気管挿管の他の方法が許容される、しかし、斜めイントロデューサーの使用は、静脈​​カテーテルの挿入のための外科的露出を最適化する少し延長位置、の首の適切な位置決めが可能。気管内カテーテルのハブは、切歯に縫合糸のループで固定し、胸部圧迫の間に頭部を固定するためにわずかな張力で保持されます。 </li><li>機械的に140μL、150回/分に設定されて齧歯類の人工呼吸器とマウスを換気する。このプロトコルは常に20〜25グラムの動物で行われるため、これは重量の調整はされません。麻酔を過剰深めるしないでください。この損なう蘇生などのエンド呼気圧力を適用するために人工呼吸器を設定しないでください。 </li><li>無菌操作や手術用顕微鏡を使用して、頸静脈に事前にフラッシュされるPE - 10カテーテルを配置。どちらの側も使用できますが、オペレータに最も近い側の使用は、胸骨圧迫や他の操作中にカテーテルdislodgementの可能性が減少することができます。 0.5％bupivicaine、0.1 mLのは、術後疼痛を制御するために、創傷の端にinfitratedています。 </li><li>シアノアクリレート系外科用接着剤で皮膚の閉鎖にPE - 10カテーテルを固定します。 </li><li>皮下心電図の電極を配置し、監視装置に接続します。信号自体の信号経路と最大化に細心の注意を払うことが蘇生の成功に不可欠です。すべてのワイヤーが動作して表面に固定されていることを確認し、信号の交差を最小限に抑え、信号経路内に絶縁体を（中空針または撚り線で空気のような）最小限に抑えることができます。 nonstrandedリード線に接続されている固体針を購入または実験室で行うことができる。接続が完了すると、モニター上にEKG信号を最適化する。 </li></ol> 2。心停止<ol><li> 37.5℃ - 直腸温36.5 ° Cとして定義され、マウスが正常体温になっていることを確認静脈内、室温0.5 M塩化カリウムの40μLを管理し、心電図上のトレース等電点電気泳動を観察。体重の用量調整はレンジ内で20から25 gの必要はありません。逮捕のタイマーを開始します。 </li><li>人工呼吸器を外します。麻酔蒸気を中止してください。加熱パッドと心電図のモニタリングを妨げる可能性のある電子的なノイズを生成する任意の他の機器の電源を切ります。マウスオーバー絶縁毛布を置きます。 </li><li>毎分は、心停止時の温度を記録します。必要に応じて、加熱ランプは、正常体温の範囲にコアの温度を起動するために使用されることがあります。心停止時には、蘇生のために物資や機器（例えば、エピネフリンの注射器）を準備する。チェックリストは、生存に必須である途切れない蘇生を、確保するために役立ちます。 </li><li> 7分、心停止の30秒後に、人工呼吸器を再接続し、事前に逮捕の設定で一回換気量を維持し、180回/分に速度を増加させる。 </li><li> 8分で、300 BPMで胸骨圧迫を開始する。心電図上のモーションアーチファクトは、裁判官CPR率に使用することができます。胸骨圧迫は、人差し指と正中線の左に剣状突起とわずかに上に5 mmを納入してください。胸は、前後の距離の1/3-1/2を圧縮する必要があり、完全なリコイルは胸骨圧迫の間で許可されている必要があります。指の位置と最適な圧縮圧力は絶対に重要です。このモデルの生存率を達成するために失敗は、ほとんど常に次善のCPRによるものです。 </li><li> CPRの最初の30秒で、15μg/ mLのに希釈エピネフリンの0.5 mLを、煎じる。合計エピネフリンの投与量は8-12μgのです。慎重に自発的な循環（ROSC）のリターンのために心電図を観察。狭い複雑なQRS群は、圧縮アーチファクトとの間で見られている。 ROSCは通常、CPRを開始した後90秒と2分の間で発生します。 ROSCは3分で発生しない場合蘇生は放棄されます。頻繁に心室性期外収縮とEKG軸の変化は、ROSC後の最初の2分で観察され、ほとんどの場合2分で安定した洞性頻脈に解決されています。 </li><li>蘇生とエピネフリンの投与量の合計時間を記録します。毎分の心拍再開後の10分間温度を記録します。 </li><li> EKGのリードは通常、ROSC後の12〜15分以内に、自発呼吸の開始時に削除することができます。 </li><li>自発呼吸数が&gt; 60/minである気管を抜管する。 </li><li>食べ物と水への容易なアクセスを確保する、℃で最初の2時間の処置後、37に設定されて温度制御された表面上回復ケージにマウスを置きます。マウスが完全に麻酔から回復し、アクティブになっているときにケージは、標準的な術後の住宅の条件に移動させることができる。 </li></ol> 3。瀉血/灌流-固定と腎臓の収穫<ol><li> CA / CPR後24時間は、イソフルラン3から4パーセントで麻酔を誘導し、ホルマリンでの使用に適しているヒュームフード内の外科表面上に仰臥位での動物を確保する.. </li><li>徐々に5％に麻酔薬の蒸気濃度を増加させることにより麻酔を深め、自発呼吸の停止によって証明されるようにマウスが深く麻酔であることを確認してください。 </li><li>クラムシェルの開胸を行い、標準的な技術に応じて心臓の左心室の心尖部穿刺によりマウスを放血させる。 </li><li>同じ針を介して、ゆっくりと注入して0.9％生理食塩水を投与する。ニックネームはさみで右心耳、灌流/瀉血をacclerateする。開腹を行い、腎臓を観察。彼らはブランチングされている場合、生理食塩水は固定用ホルマリン4％に変更される場合があります。腎臓は、尿細管上皮壊死の定量化のためのフッ素ヒスイのBとパラフィンとステンドグラスに埋め込まれています。 </li></ol> 4。代表的な結果： CAが誘導されると、動脈圧（MAP）とゼロに近いの地域の腎血流量（RRBCF）ドロップを意味し、蘇生を開始するまで（ 図1）安定している。 CA / CPR後24時間、腎機能の血清指標（血中尿素窒素、BUNおよびクレアチニン）が有意に偽手術動物と比較して上昇している。 AST / ALTは、深遠な全身の虚血（ 図2）の証拠を提供し、また上昇です。好中球ゼラチナーゼ関連リポカリン（NGAL）、腎血性傷害の感度の高い指標は、CA / CPR（ 図3）の後に大規模な上方制御24時間です。最後に、顕微鏡写真は、ヘマトキシリンおよびエオシン染色（ 図4、パネルA）とフッ素ヒスイのBで染色した髄質尿細管上皮の広範な細胞死に充填尿細管上皮および管腔の菲薄化と虚血性腎傷害の典型的な斑状、髄質壊死を示す（ 図4、パネルB）。 <img src="/files/ftp_upload/3116/3116fig1.jpg" alt="図1" /> 図1心停止は心停止の期間を通じて、地域腎皮質血流量のほぼ完全な停止（RRCBF）で、その結果大腿動脈で測定された動脈圧（MAP）は、（影付きの平均としてここに示して、灌流圧のインスタント損失を誘発する面積）。胸骨圧迫とエピネフリンを返しますMAP正常にし、RRCBFと蘇生は着実にポスト蘇生期間に上昇する。14から許可を得て転載 。 <img src="/files/ftp_upload/3116/3116fig2.jpg" alt="図2" /> 図2 24時間処置後、血中尿素窒素（BUN）、血清クレアチニン、および尿細管細胞死の程度は、すべてのような偽の手順で処理した動物と比較してCA / CPRを受けた有する動物で有意に上昇です。偽処置動物と比較して、CA / CPRは、ここでCA / CPRマウスに肝機能酵素の大規模な上昇（ALT）アラニンアミノトランスフェラーゼとアスパラギン酸アミノトランスフェラーゼ（AST）によって証明される汎organismal虚血性傷害を、引き起こす。 <img src="/files/ftp_upload/3116/3116fig3.jpg" alt="図3" /> 図3。ウエスタンブロットでは、好中球ゼラチナーゼ関連リポカリン（NGAL）、腎血性傷害の敏感な指標にポリクローナル抗体を使用して実行。尿サンプルは、（上記のA、B、C、およびDというラベルが付いています）すぐに前に（&quot;プレ&quot;）と4匹のCA / CPR（&quot;24&quot;）の後に24時間を得られた。 NGALは、大規模マウスにおいてアップレギュレートされCA / CPR後の尿は14から許可を得て転載 。 <img src="/files/ftp_upload/3116/3116fig4.jpg" alt="図4" /> 図4。）ヘマ ​​トキシリンとエオシンでは、CA / CPR後の腎組織の24の短軸の肺門部の汚れ。管状プラグと延髄とcorticomedullary細管への斑状が明確なダメージがあります。矢印はcorticomedullary接合部の腫れ、濃縮した核と、破損した細管を指す。 B）Flouro -ヒスイBは、CA / CPRの後に、同じ動物で同じ地域の24を染色。フッ素ヒスイBの染色壊死細胞明るい緑、斑状corticomedullary尿細管壊死を示す。矢印はcorticomedullary接合部で明るくステンド損傷した細管を指す。これらの知見は、アキを開発し、AKIの他の動物モデルで生成されるものとは違って、人間から腎生検所見と実質的に類似しています。

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利害の衝突は宣言されません。

マウスでの心停止と心肺蘇生の正常体温モデルはAKI、全身の血流の低下の最も一般的な臨床的な原因の病態生理および形態を複製するモデルでの評価の複数の手段を提供します。仮説検定は、遺伝子操作技術と実験用マウスのよく理解され特徴づけ解剖学や生理学の盛装へのアクセスによって促進されることがあります。 ここで説明したように、経験豊富な手の中に生?...

<table border="1"><tbody><tr><th>試薬の名前</th><th>会社</th><th>カタログ番号</th></tr><tr><td>塩化カリウム</td><td>アボットメディカル</td><td> 06653〜5 </td></tr><tr><td>イソフルレン</td><td>アボットメディカル</td><td> 05260〜05（およびその他） </td></tr><tr><td>エピネフリン塩酸</td><td>複数の</td><td>複数の</td></tr><tr><td> Digi - Keyはセンスの温度コントローラ</td><td>コー​​ル - パーマー</td><td> EW - 89000〜00 </td></tr><tr><td> Angiocath IVキャス</td><td> VWR </td><td> 381134 </td></tr><tr><td> Frova角度気道イントロデューサー</td><td>クック</td><td> G27282 </td></tr><tr><td>マウス用MicroVentベンチレータ</td><td>ハーバード装置</td><td> 733591 </td></tr></tbody></table>

normothermic cardiac arrest and cardiopulmonary resuscitation: a mouse model of ischemia-reperfusion injury

急性腎障害（AKI）は、高い死亡率1-4および最も頻繁に全身の血流の低下によって引き起こされている重大な病気の一般的な、非常に致死、合併症です。齧歯類モデルにおける全身の血流低下の5,6成功再現は、 7-9,9,10局所的虚血を用いるモデルを繰り返し臨床設定に変換されない結果を実証した、と遺伝子操作の完全なツールセットに全身の血流低下の欠如のアクセスを可能にする大規模な動物モデルを困難に満ちている事を。持っていますマウスで可能。11,12しかし、近年では心停止と心肺蘇生のマウスモデルは、モデルAKIに適応することができる浮上している。13はこのモデルは、臨床AKIに見られるように、生理的機能、解剖学的、および組織学的成果を確実に再現、急速に反復可能であり、そして遺伝子操作的手法、大動物への低コストの相対的な、そして使いやすさへのアクセスを含むマウス外科モデルの大きな利点のすべてを、提供しています。当社グループは、AKIの臓器特異的な結果の数を評価するために、このモデルの使用と豊富な経験を積んできました。14,15

Watch this Scientific Journal Video about Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury at JoVE.com

Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

周術期およびクリティカルケア関連の急性腎障害のための強力なモデルが提示される。それはほとんど臨床AKIの組織学的および機能的な変更を複製することが可能である心停止により誘発される全身の血流低下を使用する。

正常体温心停止と心肺蘇生法：虚血再灌流障害のマウスモデル

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused ...

normothermic-cardiac-arrest-cardiopulmonary-resuscitation-mouse-model

Research

JoVE Journal

Medicine

17.9K ビュー.  Oregon Health & Sciences University. 周術期およびクリティカルケア関連の急性腎障害のための強力なモデルが提示される。それはほとんど臨床AKIの組織学的および機能的な変更を複製することが可能である心停止により誘発される全身の血流低下を使用する。

ビデオ: Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often unreported mortality rates that may confound analyses. Bilateral renal pedicle clamping is commonly used to induce IR-AKI, but differences between effective clamp pressures and/or renal responses to ischemia between kidneys often lead to more variable results. In addition, shorter clamp times are known to induce more variable tubular injury, and while mice undergoing bilateral injury with longer clamp times develop more consistent tubular injury, they often die within the first 3 days after injury due to severe renal insufficiency. To improve post-injury survival and obtain more consistent and predictable results, we have developed two models of unilateral ischemia-reperfusion injury followed by contralateral nephrectomy. Both surgeries are performed using a dorsal approach, reducing surgical stress resulting from ventral laparotomy, commonly used for mouse IR-AKI surgeries. For induction of moderate injury BALB/c mice undergo unilateral clamping of the renal pedicle for 26 min and also undergo simultaneous contralateral nephrectomy. Using this approach, 50-60% of mice develop moderate AKI 24 hr after injury but 90-100% of mice survive. To induce more severe AKI, BALB/c mice undergo renal pedicle clamping for 30 min followed by contralateral nephrectomy 8 days after injury. This allows functional assessment of renal recovery after injury with 90-100% survival. Early post-injury tubular damage as well as post injury fibrosis are highly consistent using this model.

We describe models of moderate and severe ischemia-reperfusion-induced kidney injury in which the mice undergo unilateral renal pedicle clamping followed by simultaneous or delayed contralateral nephrectomy, respectively. These models consistently give rise to renal dysfunction and post-injury fibrosis, but injury severity and survival are dependent on mouse background, age and surgical equipment.

マウスにおける急性腎障害とポスト傷害線維の虚血再灌流モデル

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

医学

Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for myocardial infarction and cardiac complications of thoracic surgery. The ability of certain drugs to precondition the myocardium against ischemia reperfusion injury has led to multiple clinical trials, with little success. The isolated heart model allows acute observation of the functional effects of ischemia reperfusion injury in real time, including the effects of various pharmacological interventions administered at any time-point before or within the ischemia-reperfusion injury window. Since brief periods of ischemia can precondition the heart against ischemic injury, in situ aortic cannulation is performed to allow for functional assessment of non-preconditioned myocardium. A saline filled balloon is placed into the left ventricle to allow for real-time measurement of pressure generation. Ischemic injury is simulated by the cessation of perfusion buffer flow, followed by reperfusion. The duration of both ischemia and reperfusion can be modulated to examine biochemical events at any given time-point. Although the Langendorff isolated heart model does not allow for the consideration of systemic events affecting ischemia and reperfusion, it is an excellent model for the examination of acute functional and biochemical events within the window of ischemia reperfusion injury as well as the effect of pharmacological intervention on cardiac pre- and postconditioning. The goal of this protocol is to demonstrate how to perform in situ aortic cannulation and heart excision followed by ischemia/reperfusion injury in the Langendorff model.

The isolated rat heart is an enduring model for ischemia reperfusion injury. Here, we describe the process of harvesting the beating heart from a rat via in situ aortic cannulation, Langendorff perfusion of the heart, simulated ischemia-reperfusion injury, and infarct staining to confirm the extent of ischemic insult.

ランゲンドルフ灌流ラット心臓における虚血再灌流傷害の誘導と評価

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for ...

Murine Model of Intestinal Ischemia-reperfusion Injury

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, hypotension, necrotizing enterocolitis, bowel transplantation, trauma and chronic inflammation. Intestinal ischemia-reperfusion (IR) injury is a consequence of acute mesenteric ischemia, caused by inadequate blood flow through the mesenteric vessels, resulting in intestinal damage. Reperfusion following ischemia can further exacerbate damage of the intestine. The mechanisms of IR injury are complex and poorly understood. Therefore, experimental small animal models are critical for understanding the pathophysiology of IR injury and the development of novel therapies.
Here we describe a mouse model of acute intestinal IR injury that provides reproducible injury of the small intestine without mortality. This is achieved by inducing ischemia in the region of the distal ileum by temporally occluding the peripheral and terminal collateral branches of the superior mesenteric artery for 60 min using microvascular clips. Reperfusion for 1 hr, or 2 hr after injury results in reproducible injury of the intestine examined by histological analysis. Proper position of the microvascular clips is critical for the procedure. Therefore the video clip provides a detailed visual step-by-step description of this technique. This model of intestinal IR injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Here we describe the detailed procedure of intestinal ischemia-reperfusion in mice which results in reproducible injury without mortality to encourage the standardization of this technique across the field. This model of intestinal ischemia-reperfusion injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

腸管虚血 - 再灌流傷害のマウスモデル

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

A Mouse Model of Retinal Ischemia-Reperfusion Injury Through Elevation of Intraocular Pressure

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, diabetic retinopathy, and retinal vascular occlusions. Rodent models of I/R injury are providing significant insights into mechanisms and treatment strategies for human I/R injury, especially with regard to neurodegenerative damage in the retinal neurovascular unit. Presented here is a protocol for inducing retinal I/R injury in mice through elevation of intraocular pressure (IOP). In this protocol, the ocular anterior chamber is cannulated with a needle, through which flows the drip of an elevated saline reservoir. Using this drip to raise IOP above systolic arterial blood pressure, a practitioner temporarily halts inner retinal blood flow (ischemia). When circulation is reinstated (reperfusion) by removal of the cannula, severe cellular damage ensues, resulting ultimately in retinal neurodegeneration. Recent studies demonstrate inflammation, vascular permeability, and capillary degeneration as additional elements of this model. Compared to alternative retinal I/R methodologies, such as retinal arterial ligation, retinal I/R injury by elevated IOP offers advantages in its&#160;anatomical specificity, experimental tractability, and technical accessibility, presenting itself as a valuable tool for examining neuronal pathogenesis and therapy in the retinal neurovascular unit.

This article describes a procedure for inducing retinal ischemia-reperfusion injury by elevated intraocular pressure in mice. Retinal ischemia-reperfusion injury by elevated intraocular pressure serves to model human pathologies characterized by compromised oxygen and nutrient delivery in the retina, enabling researchers to examine potential cellular mechanisms and treatments for human diseases of the retinal neurovascular unit.

眼圧の上昇を通じて、網膜虚血再灌流障害のマウスモデル

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, ...

Ultrasonographic Assessment During Cardiopulmonary Resuscitation

The US-CAB (Ultrasound, Circulation/Airway/Breathing) protocol integrates several sonographic techniques into a structured assessment of the circulation, airway, and breathing status of a patient during cardiopulmonary resuscitation (CPR) in an advanced life support-compliant manner. US-C provides a subxiphoid view of the heart, to look for potentially reversible causes of disease, such as pericardial effusion, pulmonary embolism, hypovolemia, and acute coronary thrombosis. Sonographic cardiac activity during CPR not only helps differentiate pseudo-pulseless electrical activity (PEA) from true PEA but also represents a higher chance of the return of spontaneous circulation (ROSC) and survival. Evaluation of the inferior vena cava (IVC) shows the fluid status of the patient and indicates the best methods to use for fluid resuscitation. If aortic dissection is suspected, a subxiphoid view of the aorta is suggested for identifying an intimal flap. Once intubation is done, tracheal ultrasound (US-A) at the suprasternal notch helps differentiate endotracheal intubation (one air-mucosal interface with one comet-tail) from esophageal intubation (double tract sign). Immediately following US-A, bilateral lung US (US-B) should be done to confirm proper bilateral ventilation using the lung sliding sign. In addition, US-C can be serially followed to see the dynamic changes in the cardiac chambers and IVC, or any cardiac contraction suggestive of ROSC. US-B can also detect coexisting lung or pleural pathologies without interfering with the performance of CPR. The main concern when implementing this method is maintaining high-quality CPR without delays in chest compressions when performing US-CAB. Rigorous training and continued practice are key to minimize any interruptions during resuscitation.

Here, we present a US-CAB (Ultrasound, Circulation/Airway/Breathing) protocol for use during cardiopulmonary resuscitation (CPR). US-C evaluates the subxiphoid view of the heart and inferior vena cava. After intubation, tracheal US (US-A) and lung US (US-B) help confirm endotracheal intubation and proper ventilation.

心肺蘇生中の超音波検査

The US-CAB (Ultrasound, Circulation/Airway/Breathing) protocol integrates several sonographic techniques into a structured assessment of the circulation, ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many areas of biomedical research due to its cost-effectiveness and its translation to humans. This protocol describes a method to create a preclinical free skin flap model in rats with ischemia-reperfusion injury. The described 3 cm x 6 cm rat free skin flap model is easily obtained after the placement of several vascular ligatures and the section of the vascular pedicle. Then, 8 h after the ischemic insult and completion of the microsurgical anastomosis, the free skin flap develops the tissue damage. These ischemia-reperfusion injury-related damages can be studied in this model, making it a suitable model for evaluating therapeutic agents to address this pathophysiological process. Furthermore, two main monitoring techniques are described in the protocol for the assessment of this animal model: transit-time ultrasound technology and laser speckle contrast analysis.

Here, we describe a pre-clinical animal model for studying the pathophysiology of ischemia-reperfusion injury in reconstructive microsurgery. This free skin flap model based on the superficial caudal epigastric vessels in the rat may also allow for the evaluation of different therapies and compounds to counteract ischemia-reperfusion injury-related damage.

再建微小外科における虚血再灌流損傷の研究のための前臨床ラットモデル

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

Delayed Intramyocardial Delivery of Stem Cells after Ischemia Reperfusion Injury in a Murine Model

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, cardiac stem cell therapy is studied by delivering SCs concurrently with the induction of myocardial injury. However, this approach presents two significant limitations: the early hostile pro-inflammatory ischemic environment may affect the survival of transplanted SCs, and it does not represent the subacute infarction scenario where SCs will likely be used. Here we describe a two-part series of surgical procedures for the induction of ischemia-reperfusion injury and delivery of mesenchymal stem cells (MSCs). This method of stem cell administration may allow for the longer viability and retention around damaged tissue by circumventing the initial immune response. A model of ischemia reperfusion injury was induced in mice accompanied by the delivery of mesenchymal stem cells (3.0 x 105), stably expressing the reporter gene firefly luciferase under the constitutively expressed CMV promoter, intramyocardially 7 days later. The animals were imaged via ultrasound and bioluminescent imaging for confirmation of injury and injection of cells, respectively. Importantly, there was no added complication rate when performing this two-procedure approach for SC delivery. This method of stem cell administration, collectively with the utilization of state-of-the-art reporter genes, may allow for the in vivo study of viability and retention of transplanted SCs in a situation of chronic ischemia commonly seen clinically, while also circumventing the initial pro-inflammatory response. In summary, we established a protocol for the delayed delivery of stem cells into the myocardium, which can be used as a potential new approach in promoting regeneration of the damaged tissue.

Stems cells are continuously investigated as potential treatments for individuals with myocardial damage, however, their decreased viability and retention within injured tissue can impact their long-term efficacy. In this manuscript we describe an alternative method for stem cell delivery in a murine model of ischemia reperfusion injury.

マウスモデルにおける虚血再灌流損傷後の幹細胞の筋内内出産の遅延

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, ...

A Rat Lung Transplantation Model of Warm Ischemia/Reperfusion Injury: Optimizations to Improve Outcomes

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods for choosing appropriate cuff sizes for the pulmonary vein (PV), pulmonary artery (PA), or bronchus (Br) are varied, thus making the determination of proper cuff size during rat lung transplantation an exercise of trial and error. By standardizing the cuffing technique to use the smallest effective cuff appropriate for the size of the vessel or bronchus, one can make the transplantation procedure safer, faster, and more successful. Since diameters of the PV, PA, and Br are related to the body weight of the rat, we present a strategy to choosing an appropriate size using a weight-based guide. Since lung volume is also related to body weight, we recommend that this relationship should also be considered when choosing the proper volume of air for donor lung inflation during warm ischemia as well as for the proper volume of PBS to be instilled during bronchoalveolar lavage (BAL) fluid collection. We also describe methods for 4th intercostal space dissection, wound closure, and sample collection from both the native and transplanted lobes.

Here, we present optimizations to a rat lung transplantation model that serve to improve outcomes. We provide a size guide for cuffs based on body weight, a measurement strategy to ascertain the 4th intercostal space, and methods of wound closure and BAL (bronchoalveolar lavage) fluid and tissue collection.

温虚血/再灌流損傷のラット肺移植モデル:転帰を改善するための最適化

From our experience with rat lung transplantation, we have found several areas for improvement. Information in the existing literature regarding methods ...

An Effective Mouse Model of Unilateral Renal Ischemia-Reperfusion Injury

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models are the only available resources that mimic the complexities of the IRI-associated damage encountered in vivo. This paper describes an effective mouse model of unilateral renal IRI that delivers highly reproducible data. Ischemia is induced by occluding the right renal pedicle for 30 min followed by reperfusion. In addition to the surgical procedure, a sequential overview of the expected physiological and histopathological changes following renal IRI will be provided by comparing data from seven different reperfusion times (4 h, 8 h, 16 h, 1 day, 2 days, 4 days, and 7 days). Critical data for planning experiments ahead, such as mean surgical time, average anesthetic consumption, and body weight changes over time, will be shared. This work will help researchers implement a reliable renal IRI model and select the appropriate reperfusion time that aligns with their intended investigative goals.

Renal ischemia-reperfusion injury is associated with high morbidity and mortality in hospitalized patients. Here, we present a simple and effective mouse model of unilateral renal ischemia-reperfusion injury and provide a sequential overview of representative pathological changes observed in the kidney.

片側性腎虚血再灌流傷害の有効マウスモデル

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models ...

A Piglet Perinatal Asphyxia Model to Study Cardiac Injury and Hemodynamics after Cardiac Arrest, Resuscitation, and the Return of Spontaneous Circulation

Neonatal piglets have been extensively used as translational models for perinatal asphyxia. In 2007, we adapted a well-established piglet asphyxia model by introducing cardiac arrest. This enabled us to study the impact of severe asphyxia on key outcomes, including the time taken for the return of spontaneous circulation (ROSC), as well as the effect of chest compressions according to alternative protocols for cardiopulmonary resuscitation. Due to the anatomical and physiological similarities between piglets and human neonates, piglets serve as good models in studies of cardiopulmonary resuscitation and hemodynamic monitoring. In fact, this cardiac arrest model has provided evidence for guideline development through research on resuscitation protocols, pathophysiology, biomarkers, and novel methods for hemodynamic monitoring. Notably, the incidental finding that a substantial fraction of piglets have pulseless electrical activity (PEA) during cardiac arrest may increase the applicability of the model (i.e., it may be used to study pathophysiology extending beyond the perinatal period). However, the model generation is technically challenging and requires various skill sets, dedicated personnel, and a fine balance of the measures, including the surgical protocols and the use of sedatives/analgesics, to ensure a reasonable rate of survival. In this paper, the protocol is described in detail, as well as experiences with adaptations to the protocol over the years.

This piglet model involves surgical instrumentation, asphyxiation until the cardiac arrest, resuscitation, and post-resuscitation observation. The model allows for multiple sampling per animal, and by using continuous invasive arterial blood pressure, ECG, and non-invasive cardiac output monitoring, it provides knowledge about hemodynamics and cardiac pathophysiology in perinatal asphyxia and neonatal cardiopulmonary resuscitation.

心停止・蘇生・自然循環復帰後の心損傷と血行動態を研究するための子豚周産期仮死モデル

Neonatal piglets have been extensively used as translational models for perinatal asphyxia. In 2007, we adapted a well-established piglet asphyxia model ...