この研究は、NIHの学内プログラムからの資金によってサポートされています。

I.細胞の調製<ol><li>細胞が移植されるすべての5匹のマウス用の80％コンフルエント225センチメートル3フラスコ約10％FBSを含むDMEM中でコンフルエンスまで増殖されています。 </li><li>細胞のメディアをオフに吸引し、カルシウムまたはマグネシウムなし10 mlのPBSで洗浄する。細胞からPBSを吸引除去する。 </li><li>トリプシンの3 mlで細胞をトリプシン処理、10〜15分待ってから、メディアの15mlでトリプシンを中和する。 </li><li>ピペットで50 mlコニカルにすべてのセルとメディア。二つの大きなフラスコは、1つの50 mlコニカル収まる。 </li><li> 4℃で7分間1600 rpmで細胞をスピン完全に上清を吸引除去する。冷PBS 10mlに細胞を再懸濁し、ゆっくりと上下にピペッティングで細胞をリンス。細胞ペレットをPBSで均一に混合していることを確認します。懸濁液は、曇りでなければなりません。 </li><li>この時点で細胞数をコールターカウンターを用いて行われます。細胞を4℃で7分間1600 rpmで再度遠心分離してい</li><li>セルの第2回遠心分離した後、として1mlのPBSで海賊すべてのPBSと再懸濁する。 </li><li>細胞を4℃で10分間3000rpmでPBS 1mlのでスピンされているPBSを吸引し、PBSはU87細胞を5μLあたり1×10 6に、最終濃度を調整するために追加され、他の細胞株は、最適な腫瘍の成長のための細胞の数が異なると集中が必要な場合があります。 </li><li>最終的なボリュームと濃度が得られた後、細胞をエッペンドルフチュー​​ブに氷の上に保持されます。 </li></ol> II。異種移植片<ol class="ualpha"><li>マウスの準備<ol><li>マウスは、ケタミン/キシラジン/生理食塩水の注入ミックスを使用して麻酔する。混合物は、ケタミン120 mg / kgで、16 Rompunのmg / kgおよび食塩水である。マウスは、個別に秤量し、自分の体重に基づいて、10グラムあたり0.1mlが与えられます。麻酔薬は、1 mlのシリンジIPに接続された26G針で与えられます。それは痛み応答の睡眠段階に到達するためにマウスの約15分かかります。あなたの疼痛反応を確認することができます彼らの足蹠をつまんでマウス。 </li><li>マウスが完全に麻酔した後、眼軟膏は乾燥からそれらを防ぐために、彼らの目に投与されています。 </li><li>マウスは、口のバーでノッチのフロントアッパー歯と定位のベッドの上に配置されます。ベッドはその後定位に配置されます。 </li><li>鼻ガードが頭部の動きを防ぐためにマウスの鼻の上に配​​置されており、耳のバーは、静かにマウスの外耳道に配置されています。頭蓋骨が骨折および/または呼吸からマウスを防ぐ可能性があるので、きつすぎる鼻バーや耳バーの配置のいずれかを行うことがないように注意してください。 </li><li>マウスの頭部は定位上の目盛りを使用してレベルがこのプロセスの間に有用であることを確認してください。 </li><li>準備には、マウス、アルコールとp​​rovidineヨウ素はヨウ素のアルコールと3アプリケーションの3アプリケーションの合計の交互マウスの頭の上に自由に適用されます。 70％アルコールパッドのいずれかの最終的なアプリケーションは、に適用されます頭。 </li></ol></li><li>外科的処置<ol><li>プロシージャ内のすべての機器は、操作を始める前に70％エタノールで楽器をオートクレーブまたは浸漬のいずれかによって滅菌されるべきである。 </li><li>切開が斜め後ろの頭蓋骨の左側後部に右眼の後ろの頭蓋骨の上に作られています。切開は10 mm〜15 mm程度長くなければなりません。 </li><li> Q-ティップを使用して、徐々に頭蓋骨を露出させ、頭蓋骨と皮膚の間にある膜を払拭するために切開を分離します。過度の出血のために血をブロットおよび監視するためにQ-ティップを使用しています。 </li><li>ブレグマを視覚化するために頭蓋骨の上部を公開します。ハミルトンシリンジポイントを使用して、ブレグマの上に配置します。頭蓋骨にドリルする穴の位置が正しいことは、ブレグマ（図2）の前方2mmの右側に1 mmである。無菌購入された外科医のフェルトマジックは、掘削する穴の位置をマークするために使用されています。穴の近くに配置されることに注意してくださいterior縫合線。 </li><li>次に、18G針を介してピンの花瓶や飼料に22G針のポイントを使用してマークされた位置に頭蓋骨に小さな穴をドリルダウンします。 </li><li>細胞の5μlを10μlのガラスハミルトンシリンジをロードします。ハミルトンシリンジは、それに接続されている33Gニードルに鈍い30G 10μlです。それらをピペッティングあるいは接続されている25G針で1ミリリットル注射器を使用するか、それらをロードする前に細胞を混在させるようにしてください。ハミルトンシリンジが読み込まれた後、頭蓋骨に穴でそれを並べるために腕に戻します。 </li><li>針が頭蓋骨のレベルになると、頭蓋骨の表面に針のバントエンドを下げ、それは頭蓋骨の表面下3mmの深さになるまで針を下げます。 </li><li>針が低下した後、それが脳に影響を与える針からの主要な外傷を避けるために2分間が残っているものとします。細胞を移植しても、backfloに注意して、6-8分の合計で1分あたり1μlの割合であるワットセルの最後が注入された後、すべてのセルが入って安定させるために、別の2分の場所に針を残す</li><li>脳から針を外して、Q-ティップで穴を清掃してください。各溶液中で、その順番で3倍、50％Actone、100％エタノール、PBSで針と注射器をきれいに進みます。 </li><li>頭蓋骨の皮膚は、この時点で縫合する準備ができました。 5-0使用すると、PBSは約3針で切開を閉じます。 </li><li>マウスは、熱パッドの上に目を覚ますと、必要に応じて眼軟膏を再適用することができます。それは自分自身で動きを示した後マウスをケージに戻すことができます（つまり、頭を持ち上げたり、肩を動かす）。 </li></ol></li></ol> III。放射線と薬物によってICインプラントの治療<ol class="ualpha"><li>放射線治療<ol><li>細胞株に応じて、マウスの治療は、5から7日後にインプラントを行うことができる。私たちの部門は、放射線治療のため常用電圧放射線治療装置を使用していますメンター·。 </li><li>そのようなBLI（生物発光イメージング）、MRIやCTなどのイメージングは​​、ランダム化前（図3A）に腫瘍を視覚化するために使用することができます。 </li><li>マウスは、以前のようにケタミン/キシラジン/生理食塩水カクテルで麻酔し、眼軟膏は、彼らの目に適用されます。 </li><li>マウスは、マウスの頭が中心に向かって指しているパイの車輪のように設定されているカスタムメイドの放射線治具に配置されています。マウスの体は鉛で放射線から遮蔽されています。 </li><li>薬物治療は、別の配送ルート、すなわちSQ、IPおよびIVを通しての前または放射線のいずれか指定することができます。 </li></ol></li></ol> IV。結果マウスは麻酔注射後45-60分以内に手続きからウェイクアップする必要があります。手順と時間が1を超えた後、マウスが難しく目覚めている場合は½時間を手続きした後、マウスが回復しない可能性があり、安楽死は、効果的な代替手段かもしれません。目の場合EREは、過度の出血であるか、またはプロシージャの安楽死の間の任意の時点で割れた頭蓋骨になるにはお勧めします。すべてがうまくいくと細胞株に依存している場合の手順が行われた後、腫瘍は2-4週間以内に形成開始する必要があります。頭蓋内異種移植モデルを使用する場合は、治療効果は、通常、生存着床後5として測定されます。これはしかし、ICモデルの兆候および症状には、死の前に発生します。マウスが示すことができる物理的な徴候のいくつかは、猫背姿勢、体重の減少、不活発である。神経学的徴候のいくつかは、発作、頭部の傾き、バランスの損失かもしれません。図3Bは、彼は死亡時に腫瘍の染色を示しています。 Kaplan-Meier法のプロットは、データ（図4）を評価するために使用されます。 50％の生存日数を算出し、コントロールに比較されます。併用群は、個々の放射線や薬物武器の追加よりも長く住んでいた場合の組み合わせが成功したされています。 <img alT = &quot;図1&quot; SRC = &quot;/ files/ftp_upload/3397/3397fig1.jpg&quot; /&gt; 図1。Stoelting定位装置。 <img alt="図2" src="/files/ftp_upload/3397/3397fig2.jpg" /> 図2マウスの頭蓋骨にドリル穴の位置の図を示します。 <img alt="図3A" src="/files/ftp_upload/3397/3397fig3A.jpg" /> GBM注入後の図3A。BLIイメージング。 <img alt="図3B" src="/files/ftp_upload/3397/3397fig3B.jpg" /> 死亡率ではGBM腫瘍の図3B。HとE。 <img alt="図4" src="/files/ftp_upload/3397/3397fig4.jpg" /> 図4。PARP阻害剤7日後の手術で注入されたU87 ICインプラントの結果。

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我々は、開示することは何もありません。

このような多形性膠芽腫などの悪性神経膠腫は、最も一般的な原発性脳腫瘍を表しており、予後不良4を持っています。 GBMの影響を受けた患者の生存率は手術、放射線、化学療法5の進歩にもかかわらず（すなわち、9〜12ヶ月後の診断）過去10年間ほぼ横ばいで推移している。神経膠腫の治療の研究のための適切なモデルの特徴は、比較的離散的な腫瘍として再現性腫瘍の位置?...

<table border="1"><tbody><tr><td>ラボ標準定位Instrutment </td><td>マウスおよびラットアダプターをStoelting </td></tr><tr><td> Betadine </td><td> 100パーセントエタノール</td></tr><tr><td> 70パーセントエタノール</td><td> 50％のアセトン</td></tr><tr><td> 6 &quot;Q-のヒント</td><td>アルコール綿棒</td></tr><tr><td>ハミルトンシリンジ80008 </td><td>エチコン5から0 PD </td></tr><tr><td> 1701 SN 31-33G / 1 &quot;/ PT3針</td><td> 1 mlのシリンジ</td></tr><tr><td> 25G針</td><td>加温パッド</td></tr><tr><td>人工涙液軟膏</td><td>非常に微細な先端が永久的なマーカー</td></tr></tbody></table>

combination radiotherapy in an orthotopic mouse brain tumor model

多形性膠芽腫（GBM）は1、最も一般的で積極的な成人の原発性脳腫瘍である。近年、そこに腫瘍浸潤のメカニズムの理解にかなりの進展が、腫瘍の直接の脳内接種では、生理学的に適切な環境2で侵襲的なプロセスを観察する機会を提供しています。限り人間の脳の腫瘍が懸念されるとして、現在利用可能な同所性モデルは、複雑な開頭手術から3腫瘍の固体片の細胞懸濁液または注入の定位注射のいずれかによって確立されています。我々の技術では、脳に直接注入するのに用いられた細胞懸濁液を作成するために組織培養から細胞を収穫します。手術時間は約30分で、マウスが一定の外科的平面にする必要があるため、注射麻酔薬が使用されます。マウスがStoetling（図1）によって行われた定位ジグ内に配置されます。船尾にER外科領域を洗浄し、準備し、切開が行われている、そしてブレグマは、開頭の位置を決定するために位置しています。開頭の位置が右に2ミリメートルとブレグマへ吻側1mmである。深さは、頭蓋骨の表面から3 mmであり、細胞は2μlを2分ごとの速度で注入されています。皮膚が5から0 PDS縫合され、マウスは、加熱パッド上で目を覚ますために許可されています。我々の経験から、細胞株に応じて、治療は手術後7〜10日から行われることができます。薬物送達は、薬剤の組成に依存しています。放射線治療のためにマウスを麻酔しており、治具カスタムメイドに入れられます。鉛は、マウスの体をカバーし、マウスの脳だけを公開しています。 GBMの腫瘍形成と新しい治療法の評価の研究は、正確で再現性脳腫瘍動物モデルが必要になります。したがって、私たちはeffectivenをテストするために、脳や腫瘍の微小環境の相互作用を研究するために、この同所脳モデ​​ルを使用して、放射線とせずに別の治療薬のESS。

Watch this Scientific Journal Video about Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model at JoVE.com

Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model

この記事の目的は、放射線研究のための同所性神経膠芽腫モデルの使用を記述することです。この資料では、注入、セル処理かかわらず移動して、頭蓋モデルを用いてマウスの放射線治療になります。

同所性マウス脳腫瘍モデルのコンビネーション放射線治療

Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the ...

combination-radiotherapy-in-an-orthotopic-mouse-brain-tumor-model

Research

JoVE Journal

Medicine

16.2K ビュー.  National Cancer Institute. この記事の目的は、放射線研究のための同所性神経膠芽腫モデルの使用を記述することです。この資料では、注入、セル処理かかわらず移動して、頭蓋モデルを用いてマウスの放射線治療になります。

ビデオ: Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model

An Orthotopic Model of Murine Bladder Cancer

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and subsequent cell adhesion. After their bladders are transurethrally catheterized and drained, animals are again catheterized to permit insertion of a platinum wire into bladders without damaging the urethra or bladder. The catheters are made of Teflon to serve as an insulator for the wire, which will conduct electrical current into the bladder to create a burn injury. An electrocautery unit is used to deliver 2.5W to the exposed end of the wire, burning away extracellular layers and providing attachment sites for carcinoma cells that are delivered in suspension to the bladder through a subsequent catheterization. Cells remain in the bladder for 90 minutes, after which the catheters are removed and the bladders allowed to drain naturally. The development of tumor is monitored via ultrasound. Specific attention is paid to the catheterization technique in the accompanying video.

Bladder tumors are established in female mice in a minimally invasive fashion through catheterization, local cauterization, and subsequent adhesion of carcinoma cells to the burn sites.

マウス膀胱癌の同所性モデル

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and ...

医学

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient management strategies. Orthotopic mouse models of oral cancer have been developed to facilitate the study of factors that impact invasion and serve as model system for evaluating anti-tumor therapeutics. In these systems, visualization of disseminated tumor cells within oral cavity tissues has typically been conducted by either conventional histology or with in vivo bioluminescent methods. A primary drawback of these techniques is the inherent inability to accurately visualize and quantify early tumor cell invasion arising from the primary site in three dimensions. Here we describe a protocol that combines an established model for squamous cell carcinoma of the tongue (SCOT) with two-photon imaging to allow multi-vectorial visualization of lingual tumor spread. The OSC-19 head and neck tumor cell line was stably engineered to express the F-actin binding peptide LifeAct fused to the mCherry fluorescent protein (LifeAct-mCherry). Fox1nu/nu mice injected with these cells reliably form tumors that allow the tongue to be visualized by ex-vivo application of two-photon microscopy. This technique allows for the orthotopic visualization of the tumor mass and locally invading cells in excised tongues without disruption of the regional tumor microenvironment. In addition, this system allows for the quantification of tumor cell invasion by calculating distances that invaded cells move from the primary tumor site. Overall this procedure provides an enhanced model system for analyzing factors that contribute to SCOT invasion and therapeutic treatments tailored to prevent local invasion and distant metastatic spread. This method also has the potential to be ultimately combined with other imaging modalities in an in vivo setting.

A comprehensive overview of the techniques involved in generating a mouse model of oral cancer and quantitative monitoring of tumor invasion within the tongue through multi-photon microscopy of labeled cells is presented. This system can serve as a useful platform for the molecular assessment and drug efficacy of anti-invasive compounds.

口腔扁平上皮癌の同所性マウスモデルにおける腫瘍細胞の浸潤の多光子イメージング

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There is little knowledge about in situ, in vivo, tumor hypoxia during intracranial development of malignant brain tumors because of lack of efficient means to monitor it in these deep-seated orthotopic tumors. Bioluminescence imaging (BLI), based on the detection of light emitted by living cells expressing a luciferase gene, has been rapidly adopted for cancer research, in particular, to evaluate tumor growth or tumor size changes in response to treatment in preclinical animal studies. Moreover, by expressing a reporter gene under the control of a promoter sequence, the specific gene expression can be monitored non-invasively by BLI. Under hypoxic stress, signaling responses are mediated mainly via the hypoxia inducible factor-1&alpha; (HIF-1&alpha;) to drive transcription of various genes. Therefore, we have used a HIF-1&alpha; reporter construct, 5HRE-ODD-luc, stably transfected into human breast cancer MDA-MB231 cells (MDA-MB231/5HRE-ODD-luc). In vitro HIF-1&alpha; bioluminescence assay is performed by incubating the transfected cells in a hypoxic chamber (0.1% O2) for 24 hr before BLI, while the cells in normoxia (21% O2) serve as a control. Significantly higher photon flux observed for the cells under hypoxia suggests an increased HIF-1&alpha; binding to its promoter (HRE elements), as compared to those in normoxia. Cells are injected directly into the mouse brain to establish a breast cancer brain metastasis model. In vivo bioluminescence imaging of tumor hypoxia dynamics is initiated 2 wks after implantation and repeated once a week. BLI reveals increasing light signals from the brain as the tumor progresses, indicating increased intracranial tumor hypoxia. Histological and immunohistochemical studies are used to confirm the in vivo imaging results. Here, we will introduce approaches of in vitro HIF-1&alpha; bioluminescence assay, surgical establishment of a breast cancer brain metastasis in a nude mouse and application of in vivo bioluminescence imaging to monitor intracranial tumor hypoxia.

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model

Bioluminescence imaging of hypoxia inducible factor-1&alpha; activity is applied to monitor intracranial tumor hypoxia development in a breast cancer brain metastasis mouse model.

マウスモデルにおける乳がんの脳転移の腫瘍の低酸素動態の in vivo生物発光イメージングに

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There ...

An Orthotopic Bladder Tumor Model and the Evaluation of Intravesical saRNA Treatment

We present a novel method for treating bladder cancer with intravesically delivered small activating RNA (saRNA) in an orthotopic xenograft mouse bladder tumor model. The mouse model is established by urethral catheterization under inhaled general anesthetic. Chemical burn is then introduced to the bladder mucosa using intravesical silver nitrate solution to disrupt the bladder glycosaminoglycan layer and allows cells to attach. Following several washes with sterile water, human bladder cancer KU-7-luc2-GFP cells are instilled through the catheter into the bladder to dwell for 2 hours. Subsequent growth of bladder tumors is confirmed and monitored by in vivo bladder ultrasound and bioluminescent imaging. The tumors are then treated intravesically with saRNA formulated in lipid nanoparticles (LNPs). Tumor growth is monitored with ultrasound and bioluminescence. All steps of this procedure are demonstrated in the accompanying video.

Establishing an orthotopic bladder tumor model to evaluate antitumor effects of intravesically delivered saRNA and monitoring tumor growth by ultrasound and bioluminescent imaging.

同所性膀胱腫瘍モデルと膀胱内Sarnaの治療の評価

We present a novel method for treating bladder cancer with intravesically delivered small activating RNA (saRNA) in an orthotopic xenograft mouse bladder ...

An Orthotopic Mouse Model of Anaplastic Thyroid Carcinoma

Several types of animal models of human thyroid carcinomas have been established, including subcutaneous xenograft and orthotopic implantation of cancer cells into immunodeficient mice. Subcutaneous xenograft models have been valuable for preclinical screening and evaluation of new therapeutic treatments. There are a number of advantages to using a subcutaneous model; 1) rapid, 2) reproducible, and 3) tumor establishment, growth, and response to therapeutic agents may be monitored by visual inspection. However, substantial evidence has shed light on the short-comings of subcutaneous xenograft models1-3. For instance, medicinal treatments demonstrating curative properties in subcutaneous xenograft models often have no notable impact on the human disease. The microenvironment of the site of xenographic transplantation or injection lies at the heart of this dissimilarity.
Orthotopic tumor xenograft models provide a more biologically relevant context in which to study the disease. The advantages of implanting diseased cells or tissue into their anatomical origin equivalent within a host animal includes a suitable site for tumor-host interactions, development of disease-related metastases and the ability to examine site-specific influence on investigational therapeutic remedies. Therefore, orthotopic xenograft models harbor far more clinical value because they closely reproduce human disease. For these reasons, a number of groups have taken advantage of an orthotopic thyroid cancer model as a research tool4-7.
Here, we describe an approach that establishes an orthotopic model for the study of anaplastic thyroid carcinoma (ATC), which is highly invasive, resists treatment, and is virtually fatal in all diagnosed patients. Cultured ATC cells are prepared as a dissociated cellular suspension in a solution containing a basement membrane matrix. A small volume is slowly injected into the right thyroid gland. Overall appearance and health of the mice are monitored to ensure minimal post-operative complications and to gauge pathological penetrance of the cancer. Mice are sacrificed at 4 weeks, and tissue is collected for histological analysis. Animals may be taken at later time-points to examine more advance progression of the disease. Production of this orthotopic mouse model establishes a platform that accomplishes two objectives: 1) further our understanding of ATC pathology, and 2) screen current and future therapeutic agents for efficacy in combating ATC.

Generation of an orthotopic mouse model of anaplastic thyroid carcinoma is described here. This technique employs surgical placement of human anaplastic thyroid cancer cells into the thyroid of immunodeficient mice, thus creating a more clinically relevant setting to study disease progression as well as screen innovative therapeutic interventions.

甲状腺未分化癌の同所性マウスモデル

Several types of animal models of human thyroid carcinomas have been established, including subcutaneous xenograft and orthotopic implantation of cancer ...

An Orthotopic Bladder Cancer Model for Gene Delivery Studies

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are needed. The tumor microenvironment can significantly influence tumor development and therapy response. It is therefore often desirable to grow tumor cells in the organ from which they originated. This protocol describes an orthotopic model of bladder cancer, in which MB49 murine bladder carcinoma cells are instilled into the bladder via catheterization. Successful tumor cell implantation in this model requires disruption of the protective glycosaminoglycan layer, which can be accomplished by physical or chemical means. In our protocol the bladder is treated with trypsin prior to cell instillation. Catheterization of the bladder can also be used to deliver therapeutics once the tumors are established. This protocol describes the delivery of an adenoviral construct that expresses a luciferase reporter gene. While our protocol has been optimized for short-term studies and focuses on gene delivery, the methodology of mouse bladder catheterization has broad applications.

Implantation of cancer cells into the organ of origin can serve as a useful preclinical model to evaluate novel therapies. MB49 bladder carcinoma cells can be grown within the bladder following intravesical instillation. This protocol demonstrates catheterization of the mouse bladder for the purpose of tumor implantation and adenoviral delivery.

遺伝子送達研究のための同所膀胱癌モデル

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are ...

An Orthotopic Murine Model of Human Prostate Cancer Metastasis

Our laboratory has developed a novel orthotopic implantation model of human prostate cancer (PCa). As PCa death is not due to the primary tumor, but rather the formation of distinct metastasis, the ability to effectively model this progression pre-clinically is of high value. In this model, cells are directly implanted into the ventral lobe of the prostate in Balb/c athymic mice, and allowed to progress for 4-6 weeks. At experiment termination, several distinct endpoints can be measured, such as size and molecular characterization of the primary tumor, the presence and quantification of circulating tumor cells in the blood and bone marrow, and formation of metastasis to the lung. In addition to a variety of endpoints, this model provides a picture of a cells ability to invade and escape the primary organ, enter and survive in the circulatory system, and implant and grow in a secondary site. This model has been used effectively to measure metastatic response to both changes in protein expression as well as to response to small molecule therapeutics, in a short turnaround time.

This orthotopic model of human prostate cancer allows for quantification of tumor size, circulating tumor cells, and formation of distinct metastasis to the lung. As cells must escape the primary organ, enter the blood stream, and implant into a secondary site, this model effectively recapitulates the scenario in humans.

ヒト前立腺癌転移の同所マウスモデル

Our laboratory has developed a novel orthotopic implantation model of  human prostate cancer (PCa). As PCa death is not due to the primary tumor, but  ...

An Orthotopic Glioblastoma Mouse Model Maintaining Brain Parenchymal Physical Constraints and Suitable for Intravital Two-photon Microscopy

Glioblastoma multiforme (GBM) is the most aggressive form of brain tumors with no curative treatments available to date.
Murine models of this pathology rely on the injection of a suspension of glioma cells into the brain parenchyma following incision of the dura-mater. Whereas the cells have to be injected superficially to be accessible to intravital two-photon microscopy, superficial injections fail to recapitulate the physiopathological conditions. Indeed, escaping through the injection tract most tumor cells reach the extra-dural space where they expand abnormally fast in absence of mechanical constraints from the parenchyma.
Our improvements consist not only in focally implanting a glioma spheroid rather than injecting a suspension of glioma cells in the superficial layers of the cerebral cortex but also in clogging the injection site by a cross-linked dextran gel hemi-bead that is glued to the surrounding parenchyma and sealed to dura-mater with cyanoacrylate. Altogether these measures enforce the physiological expansion and infiltration of the tumor cells inside the brain parenchyma. Craniotomy was finally closed with a glass window cemented to the skull to allow chronic imaging over weeks in absence of scar tissue development.
Taking advantage of fluorescent transgenic animals grafted with fluorescent tumor cells we have shown that the dynamics of interactions occurring between glioma cells, neurons (e.g. Thy1-CFP mice) and vasculature (highlighted by an intravenous injection of a fluorescent dye) can be visualized by intravital two-photon microscopy during the progression of the disease.
The possibility to image a tumor at microscopic resolution in a minimally compromised cerebral environment represents an improvement of current GBM animal models which should benefit the field of neuro-oncology and drug testing.

We have established a cortical orthotopic glioblastoma model in mice for intravital two-photon microscopy that recapitulates the biophysical constraints normally at play during the growth of the tumor. A chronic glass window replacing the skull above the tumor enables the follow-up of the tumor progression over time by two-photon microscopy.

同所神経膠芽腫モデルマウスは、脳実質細胞の物理的制約を維持し、生体内二光子顕微鏡に適し

Glioblastoma multiforme (GBM) is the most aggressive form of brain tumors with no curative treatments available to date.
Murine models of this pathology ...

Tumor Treating Field Therapy in Combination with Bevacizumab for the Treatment of Recurrent Glioblastoma

A novel device that employs TTF therapy has recently been developed and is currently in use for the treatment of recurrent glioblastoma (rGBM). It was FDA approved in April 2011 for the treatment of patients 22 years or older with rGBM. The device delivers alternating electric fields and is programmed to ensure maximal tumor cell kill1.
Glioblastoma is the most common type of glioma and has an estimated incidence of approximately 10,000 new cases per year in the United States alone2. This tumor is particularly resistant to treatment and is uniformly fatal especially in the recurrent setting3-5. Prior to the approval of the TTF System, the only FDA approved treatment for rGBM was bevacizumab6. Bevacizumab is a humanized monoclonal antibody targeted against the vascular endothelial growth factor (VEGF) protein that drives tumor angiogenesis7. By blocking the VEGF pathway, bevacizumab can result in a significant radiographic response (pseudoresponse), improve progression free survival and reduce corticosteroid requirements in rGBM patients8,9. Bevacizumab however failed to prolong overall survival in a recent phase III trial26. A pivotal phase III trial (EF-11) demonstrated comparable overall survival between physicians&#8217; choice chemotherapy and TTF Therapy but better quality of life were observed in the TTF arm10.
There is currently an unmet need to develop novel approaches designed to prolong overall survival and/or improve quality of life in this unfortunate patient population. One appealing approach would be to combine the two currently approved treatment modalities namely bevacizumab and TTF Therapy. These two treatments are currently approved as monotherapy11,12, but their combination has never been evaluated in a clinical trial. We have developed an approach for combining those two treatment modalities and treated 2 rGBM patients. Here we describe a detailed methodology outlining this novel treatment protocol and present representative data from one of the treated patients.

A novel methodology that is employed for the treatment of recurrent glioblastomas is described. This treatment approach employs the application of alternating electric tumor treating fields (TTFields), known as TTF Therapy in combination with bevacizumab, a targeted agent that is currently FDA approved as monotherapy.

再発神経膠芽腫の治療のためのベバシズマブと組み合わせてフィールド療法の治療腫瘍

A novel device that employs TTF therapy has recently been developed and is currently in use for the treatment of recurrent glioblastoma (rGBM). It was FDA ...

Dynamic Contrast Enhanced Magnetic Resonance Imaging of an Orthotopic Pancreatic Cancer Mouse Model

Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) has been limitedly used for orthotopic pancreatic tumor xenografts due to severe respiratory motion artifact in the abdominal area. Orthotopic tumor models offer advantages over subcutaneous ones, because those can reflect the primary tumor microenvironment affecting blood supply, neovascularization, and tumor cell invasion. We have recently established a protocol of DCE-MRI of orthotopic pancreatic tumor xenografts in mouse models by securing tumors with an orthogonally bent plastic board to prevent motion transfer from the chest region during imaging. The pressure by this board was localized on the abdominal area, and has not resulted in respiratory difficulty of the animals. This article demonstrates the detailed procedure of orthotopic pancreatic tumor modeling using small animals and DCE-MRI of the tumor xenografts. Quantification method of pharmacokinetic parameters in DCE-MRI is also introduced. The procedure described in this article will assist investigators to apply DCE-MRI for orthotopic gastrointestinal cancer mouse models.

The goal of this protocol is to apply dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) for orthotopic pancreatic tumor xenografts in mice. DCE-MRI is a non-invasive method to analyze microvasculature in a target tissue, and useful to assess vascular response in a tumor following a novel therapy.