著者らは、寛大にマウス表現型解析方法で彼女の専門知識を共有するため、彼女の継続的な技術支援のため、ならびにための国立心臓、肺、および国立衛生研究所の血液研究所（NHLBI）（NIH）のミケーレ・アレンに感謝したいと思います私たちは、遮蔽装置を開発助けるためNHLBIのティモシー・ハント。この研究は、NIHの看護研究の国立研究所の学内研究部門によってサポートされており、検証試験の一部は社会財団看護オンコロジーからの助成金によってサポートされています。

倫理に関する声明：この研究は、米国立衛生研究所（NIH）動物実験委員会によって承認されました。動物の取り扱いと研究成果の測定に参加するすべての研究者が適切に動物管理使用のNIH事務所と国立心肺血液研究所マウス表現型コアによって訓練されました。本研究で用いたすべての動物実験の側面、住宅、環境条件は実験動物の管理と使用11用のガイドを遵守していました。 1.住宅と実験動物注：ハウス実験を通して個別に（到着時にはおよそ5週齢）の雄C57BL / 6マウスと食物および水を自由摂取のアクセスを提供します。すべてのケージは、光位相が午後6時午前6時と暗期で始まると12：12時間の明暗サイクルで保持されます。 <ol><li>マウスを特定し、個々の標準換気に割り当てますDマウスケージ。回復するための識別手続き後24時間を許可します。 注：テール入れ墨が耳タグが走行輪に引っ掛かる可能性を排除するための識別の手段として推奨されています。マウスのケージに書かれた番号と一致する尾の数字で、各マウスの尾の数刺青。 </li><li> 1日3分間静かに各マウスを扱う、マウスは、少なくとも3日間以上ケージに順応することを可能にします。 </li></ol>ホイール順化およびベースラインを実行している2 <ol><li>個々のVWRAケージ、連続記録のための電子カウンタに接続されたランニングホイールを装備し、それぞれにマウスをご紹介します。 注：すべてのホイールカウンタは、単一のUSBインターフェイスを介してコンピュータに接続する（材料リストを参照してください）。コンピュータソフトウェアは、距離が指定された総duratioの各指定の時間間隔を横切って旅し、平均速度、車輪の回転数を算出し、n個。記録が停止すると、データが自動的にテキストとしてとスプレッドシートの両方として保存されます。 </li><li>コンピュータソフトウェアインタフェースを介してVWRAの記録を開始します。少なくとも5日間に1時間と持続時間に間隔を記録する設定します。少なくとも5日間VWRAを記録し続けます。 注：ステップ2.2の終わりに、すべてのマウスは、毎日の車輪走行活動の比較的一定量を達成する必要があります。そうでない場合には、任意の外れ値を識別して排除します。 </li><li> VWRAはソフトウェアインタフェースを介して記録を停止し、（車輪を実行せずにケージ）ステップ1.1で説明したそれらの標準ケージにマウスを返します。 </li><li>偽照射対照または照射群のいずれかにマウスをランダム。 </li></ol> 3.照射注：3つの連続した日間、1日に1回、両方のグループにすべてのマウスに対して、次の手順を実行します。毎日同じ順序でマウスを扱います。 <ol><li>の腹腔内注射することにより、各マウスを麻酔ケタミン（100mg / kg）およびキシラジン（10mg / kg）の混合物。 </li><li>つま先のピンチで麻酔を確認し、麻酔下ながら乾燥を防ぐために、目に軟膏を使用しています。 </li><li>鉛遮蔽装置に麻酔マウスを転送します。唯一の下腹部/骨盤領域が露出するようにシールドにマウスを配置します。 注：シールドは、それはマウスの小、標的領域への放射線被曝を可能にするの間の狭いオープンスペースで「ボックス」2本のリードで構成されている（ 図1参照）。 </li><li>シールド内の位置でマウスの尾の付け根を確保するために医療用テープを使用してください。 注：ステップ3.4はオプションですが、そうすることが、マウスの位置は、次のステップ中に変化しないことを確実にするのに役立つことができます。 </li><li>シールド内の動物の位置が維持されることを確認して、照射に遮蔽装置を運びます。 </li><li>マウスは照射群である場合、DOSで800センチグレイをお届け約110センチグレイ/分の電子速度。マウスは、偽照射対照群である場合には、等価時間のために非アクティブ照射でマウスを残します。 注：最適な照射装置の設定は、特定のデバイスに依存します。 137セシウム源から送ら110センチグレイ/分の線量率は、ここで使用される照射の中央の線量率です。露光時間は800 cGyで、所望の総用量に達するように調整しました。 </li><li>照射とシールドからマウスを外し、ステップ1.1で説明した元の、標準のケージに戻します。 </li><li>それは胸骨横臥位を維持するのに十分な意識を取り戻したまで継続的にマウスを監視します。 </li></ol> 4.放射線誘発疲労測定<ol><li>照射の3日連続の完了後に日に、ステップ2.1で説明した個々のVWRAケージにマウスを転送します。 </li><li>ここで録音設定を除いて、ステップ2.2で説明したように録音VWRA、15日以上の期間。 15日の終わりには、手動でソフトウェアインタフェースを介してVWRA記録を停止します。 注：各記録期間からのデータは自動的に記録の期間を通して、すべての動物（列）と、すべての間隔（行）のための回転、距離、速度の測定値をそれぞれ含むスプレッドシートとして保存されます。前照射VWRAのための1つ、および照射後VWRAための1：実験の終了時に、記録ソフトウェアによって生成された2つのスプレッドシートがあります。 </li></ol>

<ol>
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この研究は、学内研究部門、看護研究の国立研究所、国立衛生研究所によってサポートされていました。

我々は、罹患率または死亡率を混乱することなく、マウスにおけるVWRAの減少を誘導するためにターゲット周辺照射を使用してプロトコルを記載しています。重要なことは、単純な遮蔽装置は、骨盤がん患者によって受信された放射線治療を模倣し、一貫して所望の領域を標的とするために、このプロトコルで照射することができます。脳または全身照射8,9<...

がん関連疲労（CRF）は、しばしば癌治療1を受けている患者に影響を与える悲惨な、高価な状態です。疲労は、最近の活性に比例も残りによって軽減でもない、それは気分、意欲、関心、および認知2に関連する擾乱の多種多様に関連付けられています。ヘモグロビンレベルおよび種々のホルモン系の機能である場合においても、炎症およびサイトカインのレベルと相関することが多くの場合に示されているがCRFの生物学的な原因は、不明である（参照Saligan ら 3生物学のレビューのためにCRFの研究）。 動物モデルを用いて制御の研究では、この複雑な条件に関連付けられた動作と生物学を理解するために必要です。一方、腫瘍関連4または化学療法に関連した5、6脂肪igueは齧歯類モデルで研究されている、CRFの病因は、治療特異的であってもよいです。放射線療法に関連するCRFを調べるために、我々のグループは最近、照射疲労7のマウスモデルを開発しました。脳または全身照射8,9を含む既存のCRFモデルとは対照的に、このモデルは、中央駆動型の挙動の変化は、疲労のような、末梢標的照射手順によってトリガすることができる方法を探ります。 ここで説明する手順は、照射による下腹部/骨盤領域を標的とするために、鉛の遮蔽を使用して、骨盤のがん患者に投与、放射線療法をモデル化するために設計されています。しかし、鉛遮蔽または実験動物へのその配置を相対的に変更することによって、この手順は、身体の他の部分の照射をモデル化するために適合させることができます。自主ホイールランニング活動（VWRA）は疲労のようなbehavioを測定するために使用されますR;それが自発的かつ正常な動作10だから、それは他の行動と生物学的試験の同時使用を可能にしなければなりません。我々は、末梢照射明白な病的状態7を引き起こすことなく、マウスにおけるVWRAを低減するのに十分であることを見出しました。このモデルを用いた将来の実験は、免疫および他の生物学的シグナル伝達に対する末梢照射だけでなく、CRFに関連した赤字を生み出すことができる中枢神経系の下流の変化の影響を明らかに役立つことがあります。

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			<td height="42" style="height:42px;width:255px;">C57BL/6 Mice</td>
			<td style="width:135px;">Charles River</td>
			<td style="width:119px;"></td>
			<td style="width:591px;">Strain code 027 (http://www.criver.com/products-services/basic-research/find-a-model/c57bl-6n-mouse)</td>
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			<td style="width:591px;">100 mg/ml stock solution</td>
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			<td></td>
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			<td style="width:135px;">(custom made)</td>
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			<td style="width:591px;">One-inch thick lead shielding arranged as two boxes with a one-inch thick gap between them for targeted irradiation</td>
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			<td height="63" style="height:63px;width:255px;">Plexiglass shielding container</td>
			<td style="width:135px;">(custom made)</td>
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			<td style="width:591px;">Plexiglass container filled with styrofoam. Styrofoam cutouts hold the lead shielding in place.</td>
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			<td style="width:135px;">Lafayette Instrument Company</td>
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			<td>http://www.lafayetteneuroscience.com/product_detail.asp?itemid=980</td>
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			<td>http://www.lafayetteneuroscience.com/product_detail.asp?itemid=1052</td>
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			<td>http://www.lafayetteneuroscience.com/product_detail.asp?itemid=1046</td>
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a mouse model of fatigue induced by peripheral irradiation

がん関連疲労（CRF）は、しばしば放射線療法を含む癌治療を受けている患者に影響を与える悲惨な、高価な状態です。ここでは、マウスでの疲労のような挙動を誘発することを目標と周辺照射を使用する方法について説明します。適切なシールドでは、照射は、骨盤の癌を有する個人が受けた放射線治療をモデル化するための努力で、脳を温存、マウスの下腹部/骨盤領域を対象としています。私たちは、明らかな病的状態を引き起こすことなく、自発的ホイールランニング活動（VWRA）により測定したマウスにおける疲労のような挙動を誘導するのに十分な照射線量を提供します。車輪走行がマウスにおいて正常な、自発的な行動であるため、その使用は、他の行動試験または生物学的な対策にはほとんど交絡効果を持っている必要があります。したがって、車輪走行疲労の挙動および生物学的相関を理解するで実現可能な結果の尺度として使用することができます。 CRFは頻繁に共同で複雑な状態であります罹患率、およびおそらく両方癌およびその種々の処理に関連した原因があります。このホワイトペーパーで説明する方法は、疲労のような末梢性トリガーが、集中駆動型行動の開発と持続性を説明することができる生物学的ネットワークを探索するために、より一般的に、CRFの発展に貢献し、放射線誘発性の変化を調査するために有用です。

マウスの3つのバッチは、上記のプロトコルを介して実行されました。 16偽および照射20（2400 cGyで、3×800センチグレイ/日）マウスの合計がありました。照射の3日連続の後、照射群が有意に偽に比べVWRAの低下を示した（混合反復測定ANOVA：照射処理、F 1,13 = 19.233、p &lt;0.001の主な効果）。効果は最低の平均VWRA距離は放射線（ 図2A）の後、三日目に?...

Watch this Scientific Journal Video about A Mouse Model of Fatigue Induced by Peripheral Irradiation at JoVE.com

A Mouse Model of Fatigue Induced by Peripheral Irradiation

我々は、マウスでの疲労のような挙動を誘発することを目標と周辺照射を使用する方法について説明します。選択された非致死照射線量は、自発的ホイールランニング活動の一週間の削減につながります。

周辺照射による疲労のマウスモデル

Cancer-related fatigue (CRF) is a distressing and costly condition that often affects patients receiving cancer treatments, including radiation therapy. ...

a-mouse-model-of-fatigue-induced-by-peripheral-irradiation

Research

JoVE Journal

Cancer Research

7.8K ビュー.  National Institutes of Health. 我々は、マウスでの疲労のような挙動を誘発することを目標と周辺照射を使用する方法について説明します。選択された非致死照射線量は、自発的ホイールランニング活動の一週間の削減につながります。 

ビデオ: A Mouse Model of Fatigue Induced by Peripheral Irradiation

Fluorescent Orthotopic Mouse Model of Pancreatic Cancer

Pancreatic cancer remains one of the cancers for which survival has not improved substantially in the last few decades. Only 7% of diagnosed patients will survive longer than five years. In order to understand and mimic the microenvironment of pancreatic tumors, we utilized a murine orthotopic model of pancreatic cancer that allows non-invasive imaging of tumor progression in real time. Pancreatic cancer cells expressing green fluorescent protein (PANC-1 GFP) were suspended in basement membrane matrix, high concentration, (e.g., Matrigel HC) with serum-free media and then injected into the tail of the pancreas via laparotomy. The cell suspension in the high concentration basement membrane matrix becomes a gel-like substance once it reaches room temperature; therefore, it gels when it comes in contact with the pancreas, creating a seal at the injection site and preventing any cell leakage. Tumor growth and metastasis to other organs are monitored in live animals by using fluorescence. It is critical to use the appropriate filters for excitation and emission of GFP. The steps for the orthotopic implantation are detailed in this article so researchers can easily replicate the procedure in nude mice. The main steps of this protocol are preparation of the cell suspension, surgical implantation, and whole body fluorescent in vivo imaging. This orthotopic model is designed to investigate the efficacy of novel therapeutics on primary and metastatic tumors.

A procedure to implant green fluorescent protein-expressing pancreatic cancer cells (PANC-1 GFP) orthotopically into the pancreas of Balb-c Ola Hsd-Fox1nu mice to assess tumor progression and metastasis is presented here.

膵臓がんの蛍光同所性マウスモデル

Pancreatic cancer remains one of the cancers for which survival has not improved substantially in the last few decades. Only 7% of diagnosed patients will ...

がん研究

Transduction-Transplantation Mouse Model of Myeloproliferative Neoplasm

Transduction-transplantation is a quick and efficient way to model human hematologic malignancies in mice. This technique results in expression of the gene of interest in hematopoietic cells and can be used to study the gene's role in normal and/or malignant hematopoiesis. This protocol provides a detailed description on how to perform transduction-transplantation using calreticulin (CALR) mutations recently identified in myeloproliferative neoplasm (MPN) as an example. In this protocol whole bone marrow cells from 5-flurouracil (5-FU) treated donor mice are transduced with a retrovirus encoding mutant CALR and transplanted into lethally irradiated syngeneic hosts. Donor cells expressing mutant CALR are marked with green fluorescent protein (GFP). Transplanted mice develop an MPN phenotype including elevated platelets in the peripheral blood, expansion of megakaryocytes in the bone marrow, and bone marrow fibrosis. We provide a step-by-step account of how to generate retrovirus, calculate viral titer, transduce whole bone marrow cells, and transplant into irradiated recipient mice.

This manuscript provides a description of the methodology used to establish transduction-transplantation mouse models. A detailed account is given of technical errors to avoid when performing bone marrow transplants. A clear understanding should be gained of the importance of high viral titer, transfection/transduction, and irradiation.

骨髄増殖性腫瘍の形質導入 - 移植マウスモデル

Transduction-transplantation is a quick and efficient way to model human hematologic malignancies in mice. This technique results in expression of the ...

Isolation of Circulating Tumor Cells in an Orthotopic Mouse Model of Colorectal Cancer

Despite the advantages of easy applicability and cost-effectiveness, subcutaneous mouse models have severe limitations and do not accurately simulate tumor biology and tumor cell dissemination. Orthotopic mouse models have been introduced to overcome these limitations; however, such models are technically demanding, especially in hollow organs such as the large bowel. In order to produce uniform tumors which reliably grow and metastasize, standardized techniques of tumor cell preparation and injection are critical. 
We have developed an orthotopic mouse model of colorectal cancer (CRC) which develops highly uniform tumors and can be used for tumor biology studies as well as therapeutic trials. Tumor cells from either primary tumors, 2-dimensional (2D) cell lines or 3-dimensional (3D) organoids are injected into the cecum and, depending on the metastatic potential of the injected tumor cells, form highly metastatic tumors. In addition, CTCs can be found regularly. We here describe the technique of tumor cell preparation from both 2D cell lines and 3D organoids as well as primary tumor tissue, the surgical and injection techniques as well as the isolation of CTCs from the tumor-bearing mice, and present tips for troubleshooting.

We describe the establishment of orthotopic colorectal tumors via injection of tumor cells or organoids into the cecum of mice and the subsequent isolation of circulating tumor cells (CTCs) from this model.

結腸直腸癌モデルマウスにおける循環腫瘍細胞の単離

Despite the advantages of easy applicability and cost-effectiveness, subcutaneous mouse models have severe limitations and do not accurately simulate ...

A Genetically Engineered Mouse Model of Sporadic Colorectal Cancer

Despite the advantages of easy applicability and cost-effectiveness, colorectal cancer mouse models based on tumor cell injection have severe limitations and do not accurately simulate tumor biology and tumor cell dissemination. Genetically engineered mouse models have been introduced to overcome these limitations; however, such models are technically demanding, especially in large organs such as the colon in which only a single tumor is desired.
As a result, an immunocompetent, genetically engineered mouse model of colorectal cancer was developed which develops highly uniform tumors and can be used for tumor biology studies as well as therapeutic trials. Tumor development is initiated by surgical, segmental infection of the distal colon with adeno-cre virus in compound conditionally mutant mice. The tumors can be easily detected and monitored via colonoscopy. We here describe the surgical technique of segmental adeno-cre infection of the colon, the surveillance of the tumor via high-resolution colonoscopy and present the resulting colorectal tumors.

A protocol for the establishment of a genetically engineered mouse model of colorectal cancer by segmental adeno-cre infection and its surveillance via high-resolution colonoscopy is presented.

散発性結腸直腸癌の遺伝子操作マウスモデル

Despite the advantages of easy applicability and cost-effectiveness, colorectal cancer mouse models based on tumor cell injection have severe limitations ...

Tumor Engraftment in a Xenograft Mouse Model of Human Mantle Cell Lymphoma

B lymphocytes are key players in immune cell circulation and they mainly home to and reside in lymphoid organs. While normal B cells only proliferate when stimulated by T lymphocytes, oncogenic B cells survive and expand autonomously in undefined organ niches. Mantle cell lymphoma (MCL) is one such B cell disorder, where the median survival rate of patients is 4 - 5 years. This calls for the need of effective mechanisms by which the homing and engraftment of these cells are blocked in order to increase the survival and longevity of patients. Therefore, the effort to develop a xenograft mouse model to study the efficacy of MCL therapeutics by blocking the homing mechanism in vivo is of utmost importance. Development of animal recipients for human cell xenotransplantation to test early stage drugs have long been pursued, as relevant preclinical mouse models are crucial to screen new therapeutic agents. This animal model is developed to avoid human graft rejection and to establish a model for human diseases, and it may be an extremely useful tool to study disease progression of different lymphoma types and to perform preclinical testing of candidate drugs for hematologic malignancies, like MCL. We established a xenograft mouse model that will serve as an excellent resource to study and develop novel therapeutic approaches for MCL.

Mantle cell lymphoma (MCL) is a difficult to treat B cell disorder and it is equally difficult to establish a xenograft mouse model of primary MCL to study and develop therapeutics. Here, we describe the successful establishment of MCL xenografts in mice to help understand its underlying biology.

人間マントル細胞リンパ腫の異種移植マウス モデルにおける腫瘍生着

B lymphocytes are key players in immune cell circulation and they mainly home to and reside in lymphoid organs. While normal B cells only proliferate when ...

Development of a Human Preclinical Model of Osteoclastogenesis from Peripheral Blood Monocytes Co-cultured with Breast Cancer Cell Lines

The crosstalk between tumor cells and bone cells in the bone microenvironment is crucial to understanding the mechanism of bone metastasis formation. We developed an in vitro fully human preclinical model of a co-culture of breast cancer cells and monocytes undergoing differentiation towards osteoclasts. We optimized a model of osteoclastogenesis starting from a sample of peripheral blood collected from healthy donors. Peripheral blood mononuclear cells (PBMCs) were first separated by density gradient centrifugation, seeded at a high density and induced to differentiate by adding two growth factors (GFs): receptor activator of nuclear factor-&#954;B ligand (RANKL) and macrophage colony-stimulating factor (MCSF). The cells were left in culture for 14 days and then fixed and analyzed by downstream analysis. In osteolytic bone metastases, one of the effects of cancer cell arrival in bone is the induction of osteoclastogenesis. We thus challenged our model with co-cultures of breast cancer cells to study the differentiation power of cancer cells with respect to GFs. A straightforward way of studying cancer cell-osteoclast interaction is to perform indirect co-cultures based on the use of conditioned medium collected from breast cancer cell cultures and mixed with fresh medium. This mixture is then used to induce osteoclast differentiation. We also optimized a method of direct co-culture in which cancer cells and monocytes undergoing differentiation share the medium and exchange secreted factors. This is a significant improvement over the original indirect co-culture method as researchers can observe the reciprocal interactions of the two cell types and perform downstream analyses for both cancer cells and osteoclasts. This method enables us to study the effect of drugs on the metastatic bone microenvironment and to seed cell lines other than those derived from breast cancer. The model can also be used to study other diseases such as osteoporosis or other bone conditions.

This protocol describes development of an in vitro human preclinical model of osteoclastogenesis from peripheral blood monocytes cultured with breast cancer cell lines to mimic the cancer cell-osteoclast interaction. The model could be used to further our understanding of bone metastasis formation and improve therapeutic options.

破骨細胞形成乳房癌細胞と共培養末梢血単球からの人間の前臨床モデルの開発

The crosstalk between tumor cells and bone cells in the bone microenvironment is crucial to understanding the mechanism of bone metastasis formation. We ...

PET and MRI Guided Irradiation of a Glioblastoma Rat Model Using a Micro-irradiator

For decades, small animal radiation research was mostly performed using fairly crude experimental setups applying simple single-beam techniques without the ability to target a specific or well-delineated tumor volume. The delivery of radiation was achieved using fixed radiation sources or linear accelerators producing megavoltage (MV) X-rays. These devices are unable to achieve sub-millimeter precision required for small animals. Furthermore, the high doses delivered to healthy surrounding tissue hamper response assessment. To increase the translation between small animal studies and humans, our goal was to mimic the treatment of human glioblastoma in a rat model. To enable a more accurate irradiation in a preclinical setting, recently, precision image-guided small animal radiation research platforms were developed. Similar to human planning systems, treatment planning on these micro-irradiators is based on computed tomography (CT). However, low soft-tissue contrast on CT makes it very challenging to localize targets in certain tissues, such as the brain. Therefore, incorporating magnetic resonance imaging (MRI), which has excellent soft-tissue contrast compared to CT, would enable a more precise delineation of the target for irradiation. In the last decade also biological imaging techniques, such as positron emission tomography (PET) gained interest for radiation therapy treatment guidance. PET enables the visualization of e.g., glucose consumption, amino-acid transport, or hypoxia, present in the tumor. Targeting those highly proliferative or radio-resistant parts of the tumor with a higher dose could give a survival benefit. This hypothesis led to the introduction of the biological tumor volume (BTV), besides the conventional gross target volume (GTV), clinical target volume (CTV), and planned target volume (PTV).
At the preclinical imaging lab of Ghent University, a micro-irradiator, a small animal PET, and a 7 T small animal MRI are available. The goal was to incorporate MRI-guided irradiation and PET-guided sub-volume boosting in a glioblastoma rat model.

In the past, small animal irradiation was usually performed without the ability to target a well-delineated tumor volume. The goal was to mimic the treatment of human glioblastoma in rats. Using a small animal irradiation platform, we performed MRI-guided 3D conformal irradiation with PET-based sub-volume boosting in a preclinical setting.

PET や MRI 誘導照射ラットの神経膠芽腫モデルのマイクロ照射器を使用して

For decades, small animal radiation research was mostly performed using fairly crude experimental setups applying simple single-beam techniques without ...

Evaluating the Role of Mitochondrial Function in Cancer-related Fatigue

Fatigue is a common and debilitating condition that affects most cancer patients. To date, fatigue remains poorly characterized with no diagnostic test to objectively measure the severity of this condition. Here we describe an optimized method for assessing mitochondrial function of PBMCs collected from fatigued cancer patients. Using a compact extracellular flux system and sequential injection of respiratory inhibitors, we examined PBMC mitochondrial functional status by measuring basal mitochondrial respiration, spare respiratory capacity, and energy phenotype, which describes the preferred energy pathway to respond to stress. Fresh PBMCs are readily available in the clinical setting using standard phlebotomy. The entire assay described in this protocol can be completed in less than 4 hours without the involvement of complex biochemical techniques. Additionally, we describe a normalization method that is necessary for obtaining reproducible data. The simple procedure and normalization methods presented allow for repeated sample collection from the same patient and generation of reproducible data that can be compared between time points to evaluate potential treatment effects.

Our goal was to develop a practical protocol to evaluate mitochondrial dysfunction associated with fatigue in cancer patients. This innovative protocol is optimized for clinical use involving only standard phlebotomy and basic laboratory procedures.

がんに関連した疲労におけるミトコンドリア機能の役割の評価

Fatigue is a common and debilitating condition that affects most cancer patients. To date, fatigue remains poorly characterized with no diagnostic test to ...

Important Endpoints and Proliferative Markers to Assess Small Intestinal Injury and Adaptation using a Mouse Model of Chemotherapy-Induced Mucositis

Intestinal adaptation is the natural compensatory mechanism that occurs when the bowel is lost due to trauma. The adaptive responses, such as crypt cell proliferation and increased nutrient absorption, are critical in recovery, yet poorly understood. Understanding the molecular mechanism behind the adaptive responses is crucial to facilitate the identification of nutrients or drugs to enhance adaptation. Different approaches and models have been described throughout the literature, but a detailed descriptive way to essentially perform the procedures is needed to obtain reproducible data. Here, we describe a method to estimate important endpoints and proliferative markers of small intestinal injury and compensatory hyperproliferation using a model of chemotherapy-induced mucositis in mice. We demonstrate the detection of proliferating cells using a cell cycle specific marker, as well as using small intestinal weight, crypt depth, and villus height as endpoints. Some of the critical steps within the described method are the removal and weighing of the small intestine and the rather complex software system suggested for the measurement of this technique. These methods have the advantages that they are not time-consuming, and that they are cost-effective and easy to carry out and measure.

Here, we present a protocol to establish important endpoints and proliferative markers of small intestinal injury and compensatory hyperproliferation using a model of chemotherapy-induced mucositis. We demonstrate the detection of proliferating cells using a cell cycle specific marker and using small intestinal weight, crypt depth, and villus height as endpoints.

化学療法誘発性粘膜炎のマウスモデルを用いて小腸損傷と適応を評価するための重要なエンドポイントと増殖マーカー

Intestinal adaptation is the natural compensatory mechanism that occurs when the bowel is lost due to trauma. The adaptive responses, such as crypt cell ...

An Oncogenic Hepatocyte-Induced Orthotopic Mouse Model of Hepatocellular Cancer Arising in the Setting of Hepatic Inflammation and Fibrosis

The absence of a clinically relevant animal model addressing the typical immune characteristics of hepatocellular cancer (HCC) has significantly impeded elucidation of the underlying mechanisms and development of innovative immunotherapeutic strategies. To develop an ideal animal model recapitulating human HCC, immunocompetent male C57BL/6J mice first receive a carbon tetrachloride (CCl4) injection to induce liver fibrosis, then receive histologically-normal oncogenic hepatocytes from young male SV40 T antigen (TAg)-transgenic mice (MTD2) by intra-splenic (ISPL) inoculation. Androgen generated in recipient male mice at puberty initiates TAg expression under control of a liver-specific promoter. As a result, the transferred hepatocytes become cancer cells and form tumor masses in the setting of liver fibrosis/cirrhosis. This novel model mimics human HCC initiation and progression in the context of liver fibrosis/cirrhosis and reflects the most typical features of human HCC including immune dysfunction.

Here, we describe the development of a clinically relevant murine model of liver cancer recapitulating the typical immune features of hepatocellular cancer (HCC).

肝炎症と線維症の設定に起因する肝細胞癌の腫瘍性肝細胞誘発性オルコトピックマウスモデル

The absence of a clinically relevant animal model addressing the typical immune characteristics of hepatocellular cancer (HCC) has significantly impeded ...