この作品は、全米科学財団のキャリア賞（CBET - 0846395）と比較医学科、Tulane大学によって資金を供給された。

1。動物<ol><li>雌C57BL/6J、マウス（ ハツカネズミ ） は出荷前にベンダーによって17ネズミのウイルスに感染外部寄生虫、寄生虫endoparasites、および抗体の自由検証されます。マウスは実験動物の管理と利用のためのガイドに従い、Tulane大学のAAALAC認定の動物施設に収容されています1。 </li><li>具体的には、マウスは、広葉樹の寝具で個別に換気されたポリカーボネートcageswithのmicroisolationフィルタートップに収容されたグループ（4-5匹/ケージ）です。彼らはアドリブでペレット化げっ歯類の食品や水道水を提供しています。 </li><li>マウスは、7日間は、研究開始前に順応さが許可されます。 </li></ol> 2。細胞<ol><li>マウス移行上皮癌細胞株MB49は、膀胱腫瘍を生成するために使用されます。 </li><li>細胞は、10％ウシ胎児血清、100単位/ mlペニシリン、および100単位/ mlストレプトマイシンを添加したダルベッコ改変イーグル培地で培養されています。 </li><li>培養は37℃とメディアの変化と、5％CO 2、2〜3日間維持されます。 </li></ol> 3。動物同所性腫瘍モデルここで説明する膀胱腫瘍モデルの点眼ギュンターらによって記載された手順の変更です。2 <ol><li>麻酔は酸素にイソフルラン2％（MWI / VetOne、メリディアン、ID、カタログ番号501017）で誘導し、0.75から1.25パーセントに維持されている。 （複数の動物をイソフルラン濃度の上昇を必要となる、これを同時に治療することができます。）角膜乾燥を防ぐために、目には人工涙液の眼軟膏で潤滑されています。体の熱は、紙のドレープの下で加熱パッドで維持されます。 </li><li>腰の毛皮は、金属製の接地板との接触を促進するためにクリップされて、そして剃毛面積はelectroconductionゲルでコーティングされています。 </li><li>各マウスは、個別に金属の接地板上に背recumbancyに置かれ、膀胱は尿道、3 / 4インチ、店頭針IVのカテーテルが除去スタイレットで潤滑、24ゲージを使用してカテーテルをしています。 </li><li>膀胱は、尿から排出される、と白金線は、カテーテルの両端に導線が残して膀胱にカテーテルを通して挿入される - カテーテルの内腔の端の約2 mm。ワイヤーは、プラチナのために酸化に対する材料の抵抗で作られています。 </li><li> 2.5 Wの現在は、凝固に設定された電気焼灼器ユニットと線の外側部分に触れて、約0.5秒のために膀胱に適用されます。 </li><li>ワイヤーは、膀胱内の別の場所や適用別の2.5 Wの充電に移動されます。 </li><li>線は削除され、1 × 10 6細胞/ mlの濃度でMB49細胞の100μlを1ミリリットルシリンジを介して膀胱内にカテーテルを介して染み込ませています。 </li><li>シリンジは、細胞懸濁液の流出を防ぐために、カテーテルに接続されたままになり、カテーテルを挿入したマウスは、細胞が火傷のサイトに接続できるように90分間麻酔下で維持されています。 </li><li>無菌生理食塩水の100〜200μlのは、リカバリの前に、各マウスにIP注射によって投与される。 </li><li>潜伏期間の終了時にカテーテルが除去され、動物は、マウスは自分のホームケージに戻される時点で、立直り反射戻り、まで加熱されたパッドの上に回復するために許可されています。 </li></ol> 4。超音波検査<ol><li>腫瘍はマウスは腫瘍の成長を監視するために膀胱の超音波検査を受ける前に3-5日のために拡張されます。 </li><li>上記のような初期の超音波検査の場合、動物は麻酔する。マウスは、背側recumbancyに配置され、腹部の皮膚は、毛皮をクリッピングすることによって調製される。 </li><li> 24ゲージ静脈カテーテルを膀胱に尿道に挿入されます。この時点で、膀胱を排出し、尿は、さらなる分析のために収集することができます。 </li><li>無菌生理食塩水100μlの超音波検査で膀胱の視認性を向上させるためにカテーテルを介して膀胱に滴下される。 </li><li>超音波伝導ゲルを腹部に適用され、超音波検査は、膀胱壁と内腔を評価するために実行されます。 </li></ol> 5。代表的な結果： 腫瘍組織はinnoculation後3-5日表示されるようになります。 5日目で - 私たちの経験では、12匹のマウスの群で、より日常的に50％以上は、（1.0μL、V =π/ 6 · L · W · Hを使用して、0.5）膀胱内で検出可能な組織の塊を持っている。メガバイト49細胞を使用する場合は、腫瘍のサイズは増大し続け、マウスは未処理行けば安楽死は、24日目で必要となります。マウスは、無気力や悪液質になったときに安楽死が推奨されて、フリルの毛皮、ハンチング、および無応答の動作、またはときに彼らは彼らの初期の体重の20％を失うが表示されます。&lt;/ P&gt; 腫瘍は急速に成長し、発展途上腫瘍の細胞が血管新生の血液供給によって保守することができるものよりも速く増殖するので、彼らは表向きは200ミリグラム以上、3のマスに到達した後に壊死のセンターを持つことになります。一つは、また間接的におそらく発展途上腫瘍内に急速な血管新生に関連しているhematurea、のレベルを介して腫瘍の進行を監視することができます。それは一貫して未治療の腫瘍を有するマウスの尿は14日目で血液が含まれていることが指摘された。赤血球の濃度は、これらのマウスでは、時間の経過とともに増加する現れ、尿の量が減少集めた。 50 mgを超える腫瘍はまた、典型的に14日後のinnoculationによって、比較的容易に触診できる。

<ol>
	<li>Institute of Laboratory Animal Resources. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Guide for the Care and Use of Laboratory Animals. , (1996).</li><li>Gunther, J. H., Jurczok, A., Wulf, T., Brandau, S., Deinert, I., Jocham, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimizing+syngeneic+orthotopic+murine+bladder+cancer+(MB49).">Optimizing syngeneic orthotopic murine bladder cancer (MB49).</a> Cancer Res. 59, 2834-2837 (1999).</li><li>Zhang, X., Atala, A., Godbey, W. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression-targeted+gene+therapy+for+the+treatment+of+transitional+cell+carcinoma.">Expression-targeted gene therapy for the treatment of transitional cell carcinoma.</a> Cancer Gene Ther. 15, 543-552 (2008).</li></ol>

動物実験は、Tulane大学のインスティテューショナルケアと使用委員会によって定められたガイドラインおよび規制に準拠して行った。

異なる結果が予想される必要がありますが、雌マウスに同所性膀胱腫瘍を確立するのこの低侵襲法は、MB49以外の細胞型に対しても適用できます。あまり積極的な腫瘍細胞が使用されている場合、（新たな最適条件を確立するために0.5 W刻みを使用して）わずかにワット数を大きくすると、腫瘍細胞の接着に、より従順エリアをもたらすでしょう、と染み込ませ、細胞のための長い膀胱のイン?...

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<td>C57Bl/6J mice</td>
<td>&nbsp;</td>
<td>Jackson Laboratories</td>
<td>000664</td>
<td>3-5-wk-old</td>
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<td>Polycarbonate cages and ventilated rack</td>
<td>&nbsp;</td>
<td>Lab Products</td>
<td>Super Mouse 750</td>
<td>327mm x 190mm x 143mm</td>
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<td>Hardwood Bedding</td>
<td>&nbsp;</td>
<td>Harlan Teklad</td>
<td>Laboratory &#x2013;grade SaniChips</td>
<td>&nbsp;</td>
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<td>Rodent diet</td>
<td>&nbsp;</td>
<td>Harlan Teklad</td>
<td>Rodent Diet 2019</td>
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<td>&nbsp;</td>
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<td>180 Plus (Vet)</td>
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<td>&nbsp;</td>
<td>Anthony Atala, Wake Forest University Baptist Medical Center</td>
<td>N.A.</td>
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an orthotopic model of murine bladder cancer

この簡単な手順では、膀胱腫瘍は、カテーテルの使用、地元の焼灼、およびその後の細胞接着を介して雌C57マウスで確立されています。彼らの膀胱は尿道カテーテルを挿入し、排水された後、動物は再び尿道や膀胱を損傷することなく、膀胱に白金線の挿入を可能にするためにカテーテルをされています。カテーテルは、火傷を作成するために膀胱内に電流を導通されるワイヤーのための絶縁体として機能するようにテフロンで作られています。電気焼灼器ユニットは、細胞外の層を離れて燃焼し、その後のカテーテルを通して膀胱に懸濁液に配信される癌細胞のための付着部位を提供し、ワイヤの露出終わりに2.5W提供する目的で使用されます。細胞は、カテーテルを除去した後、90分間膀胱、そして自然に排出することを許可膀胱内に残ります。腫瘍の発達は、超音波を介して監視されます。特定の注意は、添付のビデオではカテーテル挿入のテクニックに支払われます。

Watch this Scientific Journal Video about An Orthotopic Model of Murine Bladder Cancer at JoVE.com

An Orthotopic Model of Murine Bladder Cancer

膀胱腫瘍は、カテーテル、局所焼灼、および火傷のサイトへの癌細胞のその後の付着を介して低侵襲的にメスのマウスで確立されている。

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 ...

an-orthotopic-model-of-murine-bladder-cancer

Research

JoVE Journal

Medicine

15.8K ビュー.  Tulane University. 膀胱腫瘍は、カテーテル、局所焼灼、および火傷のサイトへの癌細胞のその後の付着を介して低侵襲的にメスのマウスで確立されている。

ビデオ: An Orthotopic Model of Murine Bladder Cancer

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 ...

医学

Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model

Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the understanding of the mechanics of tumor invasion, and direct intracerebral inoculation of tumor provides the opportunity of observing the invasive process in a physiologically appropriate environment2. As far as human brain tumors are concerned, the orthotopic models currently available are established either by stereotaxic injection of cell suspensions or implantation of a solid piece of tumor through a complicated craniotomy procedure3. In our technique we harvest cells from tissue culture to create a cell suspension used to implant directly into the brain. The duration of the surgery is approximately 30 minutes, and as the mouse needs to be in a constant surgical plane, an injectable anesthetic is used. The mouse is placed in a stereotaxic jig made by Stoetling (figure 1). After the surgical area is cleaned and prepared, an incision is made; and the bregma is located to determine the location of the craniotomy. The location of the craniotomy is 2 mm to the right and 1 mm rostral to the bregma. The depth is 3 mm from the surface of the skull, and cells are injected at a rate of 2 &mu;l every 2 minutes. The skin is sutured with 5-0 PDS, and the mouse is allowed to wake up on a heating pad. From our experience, depending on the cell line, treatment can take place from 7-10 days after surgery. Drug delivery is dependent on the drug composition. For radiation treatment the mice are anesthetized, and put into a custom made jig. Lead covers the mouse's body and exposes only the brain of the mouse. The study of tumorigenesis and the evaluation of new therapies for GBM require accurate and reproducible brain tumor animal models. Thus we use this orthotopic brain model to study the interaction of the microenvironment of the brain and the tumor, to test the effectiveness of different therapeutic agents with and without radiation.

The purpose of this article is to describe the use of an orthotopic glioblastoma model for chemoradiation studies. This article will go though cell processing, implanting, and radiotherapy of the mouse using an intracranial model.

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 ...

Development of Obliterative Bronchiolitis in a Murine Model of Orthotopic Lung Transplantation

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and standardized not only for the study of allo-rejection but also between syngeneic strains for examining mechanisms of ischemia-reperfusion injury after lung transplantation. Although the application of the rat and other large animal model 2 contributed significantly to the elucidation of these studies, the scope of those investigations is limited by the scarcity of knockout and transgenic rats. Due to no effective therapies for obliterative bronchiolitis, the leading cause of death in lung transplant patients, there has been an intensive search for pre-clinical models that replicate obliterative bronchiolitis. The tracheal allograft model is the most widely used and may reproduce some of the histopathologic features of obliterative bronchiolitis 3. However, the lack of an intact vasculature with no connection to the recipient's conducting airways, and incomplete pathologic features of obliterative bronchiolitis limit the utility of this model 4. Unlike transplantation of other solid organs, vascularized mouse lung transplants have only recently been reported by Okazaki and colleagues for the first time in 2007 5. Applying the basic principles of the rat lung transplant, our lab initiated the obliterative bronchiolitis model using minor histoincompatible antigen murine orthotopic single-left lung transplants which allows the further study of obliterative bronchiolitis immunopathogenesis6.

Obliterative bronchiolitis is the key impediment to the long-term survival of lung transplant recipients and the lack of a robust preclinical model precludes examining obliterative bronchiolitis immunopathogenesis. Unlike other solid organ transplants, vascularized mouse lung transplantation has only recently been developed. Here we show our independently developed obliterative bronchiolitis model after murine orthotopic single-lung transplantation.

同所性肺移植のマウスモデルにおける閉塞性細気管支炎の開発

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and ...

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 ...

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules that regulate HSCs. In these transplant model systems, the function of HSCs is determined by the ability of these cells to engraft and reconstitute lethally irradiated recipient mice. Commonly, the donor cell contribution/engraftment is measured by antibodies to donor- specific cell surface proteins using flow cytometry. However, this method heavily depends on the specificity and the ability of the cell surface marker to differentiate donor-derived cells from recipient-originated cells, which may not be available for all mouse strains. Considering the various backgrounds of genetically modified mouse strains in the market, this cell surface/ flow cytometry-based method has significant limitations especially in mouse strains that lack well-defined surface markers to separate donor cells from congenic recipient cells. Here, we reported a PCR-based technique to determine donor cell engraftment/contribution in transplant recipient mice. We transplanted male donor bone marrow HSCs to lethally irradiated congenic female mice. Peripheral blood samples were collected at different time points post transplantation. Bone marrow samples were obtained at the end of the experiments. Genomic DNA was isolated and the Y chromosome specific gene, Zfy1, was amplified using quantitative Real time PCR. The engraftment of male donor-derived cells in the female recipient mice was calculated against standard curve with known percentage of male vs. female DNAs. Bcl2 was used as a reference gene to normalize the total DNA amount. Our data suggested that this approach reliably determines donor cell engraftment and provides a useful, yet simple method in measuring hematopoietic cell reconstitution in murine bone marrow transplantation models. Our method can be routinely performed in most laboratories because no costly equipment such as flow cytometry is required.

Determining donor cell engraftment presents a challenge in mouse bone marrow transplant models that lack well-defined phenotypical markers. We described a methodology to quantify male donor cell engraftment in female transplant recipient mice. This method can be used in all mouse strains for the study of HSC functions.

競争力のあるマウス骨髄移植モデルにおけるドナー細胞の生着を​​決定するために、定量的リアルタイムPCRを用いて

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%. New therapeutic options are critically needed and depend on improved understanding of pancreatic cancer biology. To better understand the interaction of cancer cells with the pancreatic microenvironment, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression. Luciferase-tagged pancreatic cancer cells are resuspended in Matrigel and delivered into the pancreatic tail during laparotomy. Matrigel solidifies at body temperature to prevent leakage of cancer cells during injection. Primary tumor growth and metastasis to distant organs are monitored following injection of the luciferase substrate luciferin, using in vivo imaging of bioluminescence emission from the cancer cells. In vivo imaging also may be used to track primary tumor recurrence after resection. This orthotopic model is suited to both syngeneic and xenograft models and may be used in pre-clinical trials to investigate the impact of novel anti-cancer therapeutics on the growth of the primary pancreatic tumor and metastasis.

Improved understanding of pancreatic cancer biology is critically needed to enable the development of better therapeutic options to treat pancreatic cancer. To address this need, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression using in vivo bioluminescence imaging.

膵臓癌の進行の生物発光同所性モデル

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%.  New therapeutic options are critically needed and depend on improved ...

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  ...

Minimally Invasive Establishment of Murine Orthotopic Bladder Xenografts

Orthotopic bladder cancer xenografts are the gold standard to study molecular cellular manipulations and new therapeutic agents&#160;in vivo. Suitable cell lines are inoculated either by intravesical instillation (model of nonmuscle invasive growth) or intramural injection into the bladder wall (model of invasive growth). Both procedures are complex and highly time-consuming. Additionally, the superficial model has its shortcomings due to the lack of cell lines that are tumorigenic following instillation. Intramural injection, on the other hand, is marred by the invasiveness of the procedure and the associated morbidity for the host mouse.
With these shortcomings in mind, we modified previous methods to develop a minimally invasive approach for creating orthotopic bladder cancer xenografts. Using ultrasound guidance we have successfully performed percutaneous inoculation of the bladder cancer cell lines UM-UC1, UM-UC3 and UM-UC13 into 50 athymic nude. We have been able to demonstrate that this approach is time efficient, precise and safe. With this technique, initially a space is created under the bladder mucosa with PBS, and tumor cells are then injected into this space in a second step. Tumor growth is monitored at regular intervals with bioluminescence imaging and ultrasound. The average tumor volumes increased steadily in in all but one of our 50 mice over the study period.
In our institution, this novel approach, which allows bladder cancer xenograft inoculation in a minimally-invasive, rapid and highly precise way, has replaced the traditional model.

The established technique to inoculate primary invasive orthotopic bladder cancer xenografts requires laparotomy and mobilization of the bladder. This procedure inflicts significant morbidity on the mice, is technically challenging and time-consuming. We therefore developed a high-precision, percutaneous approach utilizing ultrasound guidance.

マウス同所性膀胱異種移植片の最小侵襲性の確立

Orthotopic bladder cancer xenografts are the gold standard to study molecular cellular manipulations and new therapeutic agents&#160;in vivo. Suitable ...