We thank Dr. Roger Erickson for his support and assistance with the preparation of the manuscript. This work was supported by the National Cancer Institute at the National Institutes of Health through grant numbers RO1CA160079, RO1CA138642, UO1CA184966 and VA funded program project number 1P1 BX001604.

動物に関わるすべての手続きのためのプロトコルを見直し、制度的動物実験委員会（IACUC）によって承認されなければなりません。実験動物の管理と使用のために公式に承認された手順に従ってください。前立腺内注射はオープン腹部手術を必要とし、動物は、適切な外科的無菌技術は、全体の手順の間に使用されている指定された手術室で病​​原体のない環境で保たれるべきです。 移植用の細胞の調製注：研究ニーズに基づいて、いずれかの前立腺癌細胞株を使用することができます。細胞株は、供給者の指示に従って培養されます。 <ol><li>安定的にホタルルシフェラーゼ遺伝子を発現するPC3Mリュック-C6細胞株のために、最小必須培地（MEM）中の培養細胞を、10％ウシ胎児血清（FBS）、1×非必須アミノ酸、1×フレオマイシンD1及び1mMのピルビン酸ナトリウムを補充し。インキュベーターワットで細胞を維持i番目の37℃で95％空気および5％CO 2の加湿雰囲気。 PC3M-リュック-C6細胞を、UCSFの中核施設から調達されました。 </li><li>トリプシン処理により細胞を回収。リン酸緩衝生理食塩水（PBS）で1回洗浄培養プレート。細胞が剥離するまで、37℃で95％空気および5％CO 2の加湿雰囲気でインキュベーター中で10cmディッシュ2mLの0.05％トリプシンを加え、3~5分間インキュベートします。 <ol><li>凝集を回避するために、細胞を分離するために待っている間に皿を押すか、振とうすることにより細胞を攪拌しないでください。完全培地5mlに細胞を収集し、200×gで5分間スピンダウン。トリプシンを除去するためにPBSで細胞ペレットを洗浄します。 </li></ol></li><li>トリパンブルー排除アッセイにより生細胞を列挙します。 0.4％の10μlのPBS中の細胞懸濁液（重量/ v）のトリパンブルー溶液10μlを混ぜます。血球計数器または計数チャンバースライドに混合物をロードし、顕微鏡下ですぐに細胞数をカウントしたり、細胞COUでチャンバースライドを読みますNTER。 <ol><li> 10μlの培地に2.5×10 5個の細胞を含む細胞懸濁液を準備します。 10μlの基底膜のような細胞外マトリックス抽出物と細胞懸濁液を混合し、細胞を氷上に置きます。細胞懸濁液にルシフェリンを追加し（1：200μlの、株式を30mg / mlの濃度）をマウスに注入する前に。 注：これは異なる実験群間の細胞注射の整合性をチェックするために、動物の即時バイオイメージングを可能にします。細胞生存率は、剥離後に急激に減少するため、トリプシン処理した後、好ましくは30分以内に、できるだけ迅速に細胞を注入します。 </li></ol></li></ol>手術領域の調製<ol><li>無菌状態を促進整頓、消毒領域に手術を行います。 </li><li>手術前に漂白剤溶液でカウンタ/実験台を消毒します。 注：アルコールの使用は、効果（15分）を取るために必要な長い接触時間に推奨されていません。 </li><li>滅菌ドレープを使用して、吸収性パッドやタオルをきれいにし、それぞれの外科手術セッションの後、これらの材料を交換してください。使用前にすべての機器を滅菌します。好ましい方法は、蒸気オートクレーブ、ガラスビーズ滅菌、エチレンオキサイドガス、または過酸化水素蒸気滅菌です。 </li><li>外科手術を行うために無菌的に洗浄解剖顕微鏡を使用するか、経験豊富な研究者が顕微鏡なしでそれを行うことができます。 </li></ol>腫瘍細胞の3移植<ol><li>男性の6〜8週齢の免疫不全のBalb / CまたはNOD / SCIDマウスを使用してください。 注：これは、小さな動物を操作することはより困難である、より大きな動物は、腫瘍の増殖および転移のより遅い反応速度を有する傾向があります。 </li><li>動物施設の指示に従って、手術前の痛みの薬を注入します。例えば、0.1 mg / kg体重の用量で腹腔内ブプレノルフィンを用いることができます。 </li><li>酸素とWA 1-3％イソフルランでイソフルラン室にそれらを配置することによって動物を麻酔それは、動物が完全に麻酔されるまで。この時点での筋緊張のないつま先の反射がないことを確認してください。全身麻酔中に眼球乾燥症による失明を防ぐために、適切な獣医学の眼軟膏の潤滑剤を使用してください。 注：65 mg / kg体重の用量で：研究者の好ましい方法などにより動物を麻酔、ペントバルビタールナトリウムのために、グラム体重あたり0.05 mgを腹腔内またはケタミン/キシラジン溶液（17.16ミリグラム/ mlの濃度）を投与します皮下投与に使用されます。イソフルラン吸入は麻酔の好ましい方法です。眼軟膏は優しく角膜擦れることなく適用されるべきです。 </li><li>イソフルランチャンバーから動物を削除し、動物が先に進む前に、全身麻酔下にあることを確実にするために酸素中で1〜2％イソフルランの連続流とノーズコーン装置内に配置します。 <ol><li>シェービングによってマウスから髪を削除するか、または手順を開始する前に、脱毛クリームを使用しています。 注：これは、手術中に滅菌加熱パッド上にマウスを配置することが好ましいであろう。 </li></ol></li><li>仰臥位でマウスを置きます。ポビドンヨード溶液を70％エタノール綿棒続いw / wの10％と下腹部をきれいにしてください。 </li><li>細かい鉗子のペアで、2ミリメートル包皮腺の上に陰茎鞘上記の約1~2センチ、および胸郭の下、以下の約2~3センチ皮膚の領域を持ち上げます。 </li><li>最初のメスで皮膚を介して、その後、シザー（ 図1）との筋肉層を介して、長さに正中切開1cmのを確認します。 </li><li>体腔内の膀胱の位置を確認します。これは、直接切開（ 図1）の下に位置する黄淡褐色球形の器官、です。 </li><li>細かい鉗子グリップのペア膀胱と陰茎鞘に向かって体腔外に下方その後、上方に持ち上げます。これは、白い袋状の器官のペアと明確に区​​別されている2精嚢を公開します。 </li><li>ととも​​に綿棒は、それぞれの手で、精嚢、一つ一つを外部化し、体腔からそれらを引き出し、それらを途中で膀胱（ 図2）と腹部の外側表面上に伏せて横たわっていました。 </li><li>綿棒を使用して、2つの背側の前立腺ローブがはっきりと見えるように優しく、陰茎鞘に向けて、膀胱頸部付近の挿入の時点で精嚢を倒します。組織損傷（ 図3）を回避するために、湿った綿棒を使用してください。 </li><li>注射器に充填する前にマイクロピペットで細胞懸濁液を攪拌します。 </li><li>綿棒で位置に精嚢を配置しながら、顕微鏡下で背側前立腺ローブ（ 図4）に注射針を挿入します。水疱形成が識別されるまで、ゆっくりと細胞懸濁液の20μLを注入します。膨らみ水疱は、注射が正しいことを示しています。 </li><li>と注射部位に軽く針、を押して後退しながら、数秒間綿棒ホールド漏れを防止します。 </li><li>慎重に綿棒と精嚢を持ち上げ、膀胱に続く1により体腔1にそれらをバックに挿入します。この手順を実行中に内臓 &#39;を駆け巡って」は避けてください。 </li><li>体腔内に戻って臓器を配置した後、非吸収性4-0ナイロン縫合糸で皮膚の閉鎖に続いて吸収4-0クロム腸線縫合糸で中断したパターンの最初の筋層を縫合。皮膚はまた、完全に切開部を閉鎖するために一緒に引っ張られ、手術用クランプを閉じることができます。 注：マウスはまた、縫合糸と一緒に推奨され引っ掻き、したがって、傷の開口を再組織接着剤の使用につながる可能性があり、その傷、で噛む習慣があります。画像動物はすぐにすべての実験群間で同等の生物発光強度があることを保証します。 </li><li>ケージをきれいにし、加温ランプや熱の下でそれらを保つために動物を返します。パッドる。彼らは完全に麻酔から回復し、胸骨横臥位を維持するまで、常に動物を監視します。 </li></ol>動物の4モニタリング<ol><li>制度的なプロトコルに従って、実験終了まで定期的に動物を監視します。外科的な金属製のクリップを使用している場合、1〜2週間後に削除します。実験期間は、特定の研究の必要性に依存します。 注記：この実験は、マウスの前立腺に移植癌細胞の正常な確立を検査するために21日間行いました。 </li><li>動物施設の指示に基づいて、鎮痛薬を管理します。必要に応じて、例えば、0.1 mg / kg体重の用量で腹腔内ブプレノルフィンは、各8~12時間、6時間および追加用量後の第2の用量で手順の際に使用することができます。 </li><li>動物体重、摂餌量、肌の色や質感、活動と排尿や排便の頻度を監視します。動物にIMMEDを安楽死させますiately 15％を超える体重の有意な損失がある場合。 <ol><li>安楽死のために、無意識と完全narcotizationが発生するようにCO 2がゆっくりとチャンバーを入力することができ、チャンバまたはケージ容積/分10〜30％を置換する流量で非混雑ケージに加圧タンクからのCO 2をお届け死の前に。 </li><li>呼吸停止後少なくとも1分間のCO 2流れを維持し、死が前に物理的な方法を実行するに起こるように十分な時間、室内で動物を残します。 </li><li>実行断頭、頸椎脱臼または任意の他のIACUCは、化学的に動物を安楽死させた後、物理的な方法を承認しました。 注：大幅に体重減少は、多くの場合、無気力状態を示しています。腫瘍を有する動物は、実験の終了時まで腫瘍の存在を除いて良好な健康状態であるべきです。安楽死は安楽死にAVMAガイドラインと整合的であるべきであるとLISTEでなければなりません承認されたIACUCプロトコル中のd。 </li></ol></li></ol> 5.動物の非侵襲的バイオイメージング<ol><li>毎週の癌細胞、腫瘍増殖および任意の遠隔転移のコロニー形成を追跡するために非侵襲的イメージング技術を用いて動物を監視します。 注：このようなGFPイメージング、ルシフェラーゼイメージング、X線または3Dマイクロコンピュータ断層撮影（UCT）などのイメージングモダリティなどは特定の研究に基づいて使用することができます12月15日を必要としています。 </li></ol>

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著者は、彼らが競合する金融利害関係を持たないことを宣言します。

この原稿は、人間の同所前立腺癌異種移植マウスモデルを確立するための詳細な手順を説明します。このモデルは、免疫不全マウスの背部前立腺ローブにヒト前立腺癌細胞株PC3Mリュック-C6の直接注入によって設立されました。腫瘍は、実験の過程で発達させました。腫瘍増殖は、実験中に非侵襲的生物発光イメージングシステムによって毎週モニターしました。 異種移?...

前立腺癌は次の肺と気管支（28％）1の癌に、米国では男性のがん死亡（9％）の第二の最も一般的な原因です。最近のデータによれば、220、800、新たに診断された前立腺癌症例27、540 deaths 2015年1で発生すると推定されている。高度な転移性疾患のことがあるが、早期前立腺癌の5年相対生存率は&gt; 99％ですわずか28％1。高度な転移性疾患の治療のための主要な課題は、特に前立腺癌のための頻繁な部位である骨に、他の臓器に転移するために、この疾患の傾向の根底にある分子機構の理解の欠如です。したがって、高度な転移性疾患の2,3への進行に対する効果的な治療法を開発するために、これらの前立腺腫瘍の分子メイクを研究する明確な必要性があります。 前立腺腫瘍は、HIGを示します進行に明確に定義された経路のない時間生物学的異質。転移は、多くの場合、腫瘍侵襲性4の無事前の指示で発生します。この臨床的不均一性は、前立腺癌の分子多様性に起因します。これらの致死的な腫瘍の分子構成を理解することは、この疾患のためのより良い診断および治療戦略を設計するためのキーです。したがって、前立腺癌の研究は、現在、理解および転移を予防に焦点を当てています。 前臨床in vivoでのマウスモデルは、高度な転移性疾患に前立腺癌の進行の分子機構を理解するためのさまざまなオプションを提供しています。さらに、これらのモデルは、この疾患に対する新たな治療戦略の前臨床評価のために重要です。最も一般的に使用される動物モデルは、トランスジェニックマウスモデル、尾静脈注射、心臓内移植およびヒト同所性マウスモデルを含みます。トランスジェニックの研究では、時間consumiありますNGとヒトのそれとマウスにおける前立腺癌発生の相関が変動11を示しています。自発的転移のマウスモデルにおいて、細胞は、循環中に直接注入されると、それらは迅速なターンアラウンド時間を有するが、それらは原発腫瘍または転移カスケード5の初期段階を研究するために使用することができません。同所異種移植モデルは、骨転移病変、前立腺癌の転移の一般的なサイトの開発の制限があります。それにもかかわらず、人間の同所前立腺癌異種移植マウスモデルは、十分に特徴付けられており、広く原発腫瘍の発達、腫瘍や臓器微小環境、治療的介入の6のための転移性疾患および実験薬物の使用の初期段階の間のクロストークの分子事象を研究するために使用され、7,8-11。

<table><tbody><tr><td>PC3 prostate cancer cell line&amp;nbsp;</td><td>ATCC</td><td>CRL-1435</td><td></td></tr><tr><td>Minimum Essential Medium (MEM)&amp;nbsp;</td><td>GIBCO,Life Technology</td><td>11095-080</td><td></td></tr><tr><td>PBS</td><td>GIBCO,Life Technology</td><td>10010-023</td><td></td></tr><tr><td>FBS</td><td>GIBCO,Life Technology</td><td>10437-028</td><td></td></tr><tr><td>Zeocin</td><td>Invitrogen,Life Technology</td><td>R250-01</td><td></td></tr><tr><td>Trypsin&amp;nbsp;</td><td>GIBCO,Life Technology</td><td>25300-54</td><td></td></tr><tr><td>IVIS&amp;nbsp;</td><td>Xenogen-Caliper</td><td></td><td></td></tr><tr><td>Insulin Syringes (300 &amp;micro;l, 28.5 g)</td><td>Becton Dickinson</td><td>309300</td><td></td></tr><tr><td>Mice</td><td>Charles River Laboratories, Inc</td><td></td><td></td></tr><tr><td>Alcohol Swabs</td><td>MEDEquip Depot</td><td>326895 BD</td><td></td></tr><tr><td>PVP Iodine Prep Pad</td><td>MEDEquip Depot</td><td>C12400PDI</td><td></td></tr><tr><td>Surgical CatGut Chromic Suture</td><td>Demetech</td><td>CC224017F0P</td><td></td></tr><tr><td>Matrigel</td><td>Corning</td><td>354248</td><td></td></tr></tbody></table>

pre-clinical orthotopic murine model of human prostate cancer

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information about this disease. The human orthotopic prostate cancer xenograft mouse model provides a useful alternative approach for understanding the specific interactions between genetically and molecularly altered tumor cells, their organ microenvironment, and for evaluation of efficacy of therapeutic regimens. This is a well characterized model designed to study the molecular events of primary tumor development and it recapitulates the early events in the metastatic cascade prior to embolism and entry of tumor cells into the circulation. Thus it allows elucidation of molecular mechanisms underlying the initial phase of metastatic disease. In addition, this model can annotate drug targets of clinical relevance and is a valuable tool to study prostate cancer progression. In this manuscript we describe a detailed procedure to establish a human orthotopic prostate cancer xenograft mouse model.

後方前立腺葉にPC3Mリュック-C6細胞の同所移植後、マウスを毎週（ - B 図5A）、実験の過程で細胞のコロニー形成および腫瘍増殖をモニターするために生きている動物の生物発光イメージングシステムを用いて画像化しました。生物発光シグナルの定量化は、PC3M-リュック-C6細胞は正常前立腺ローブをコロニー形成することを示しました。増加?...

Watch this Scientific Journal Video about Pre-clinical Orthotopic Murine Model of Human Prostate Cancer at JoVE.com

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

Prostate cancer is the second most common cause of cancer-related deaths in the United States. An orthotopic cancer model provides a useful approach to understand the biology of prostate cancer and to evaluate the efficacy of therapeutic regimens. This protocol describes detailed steps necessary to establish an orthotopic prostate cancer mouse model.

ヒト前立腺癌の前臨床同所性マウスモデル

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

pre-clinical-orthotopic-murine-model-of-human-prostate-cancer

Research

JoVE Journal

Medicine

14.2K ビュー.  VA Medical Center and UCSF. Prostate cancer is the second most common cause of cancer-related deaths in the United States. An orthotopic cancer model provides a useful approach to understand the biology of prostate cancer and to evaluate the efficacy of therapeutic regimens. This protocol describes detailed steps necessary to establish an orthotopic prostate cancer mouse model.

ビデオ: Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

A Murine Orthotopic Bladder Tumor Model and Tumor Detection System

This protocol describes the generation of bladder tumors in female C57BL/6J mice using the murine bladder cancer cell line MB49, which has been modified to secrete human Prostate Specific Antigen (PSA), and the procedure for the confirmation of tumor implantation. In brief, mice are anesthetized using injectable drugs and are made to lay in the dorsal position. Urine is vacated from the bladder and 50 &#181;L of poly-L-lysine (PLL) is slowly instilled at a rate of 10 &#181;L/20 s using a 24 G IV catheter. It is left in the bladder for 20 min by stoppering the catheter. The catheter is removed and PLL is vacated by gentle pressure on the bladder. This is followed by instillation of the murine bladder cancer cell line (1 x 105 cells/50 &#181;L) at a rate of 10 &#181;L/20 s. The catheter is stoppered to prevent premature evacuation. After 1 h, the mice are revived with a reversal drug, and the bladder is vacated. The slow instillation rate is important, as it reduces vesico-ureteral reflux, which can cause tumors to occur in the upper urinary tract and in the kidneys. The cell line should be well re-suspended to reduce clumping of cells, as this can lead to uneven tumor sizes after implantation.
This technique induces tumors with high efficiency. Tumor growth is monitored by urinary PSA secretion. PSA marker monitoring is more reliable than ultrasound or fluorescence imaging for the detection of the presence of tumors in the bladder. Tumors in mice generally reach a maximum size that negatively impacts health by about 3 - 4 weeks if left untreated. By monitoring tumor growth, it is possible to differentiate mice that were cured from those that were not successfully implanted with tumors. With only end-point analysis, the latter may be mistakenly assumed to have been cured by therapy.

This protocol describes the generation of murine orthotopic bladder tumors in female C57BL/6J mice and the monitoring of tumor growth.

マウス同所性膀胱腫瘍モデルおよび腫瘍検出システム

This protocol describes the generation of bladder tumors in female C57BL/6J mice using the murine bladder cancer cell line MB49, which has been modified ...

がん研究

A Bioluminescent and Fluorescent Orthotopic Syngeneic Murine Model of Androgen-dependent and Castration-resistant Prostate Cancer

Orthotopic tumor modeling is a valuable tool for pre-clinical prostate cancer research, as it has multiple advantages over both subcutaneous and transgenic genetically engineered mouse models. Unlike subcutaneous tumors, orthotopic tumors contain more clinically accurate vasculature, tumor microenvironment, and responses to multiple therapies. In contrast to genetically engineered mouse models, orthotopic models can be performed with lower cost and in less time, involve the use of highly complex and heterogeneous mouse or human cancer cell lines, rather that single genetic alterations, and these cell lines can be genetically modified, such as to express imaging agents. Here, we present a protocol to surgically injecting a luciferase- and mCherry-expressing murine prostate cancer cell line into the anterior prostate lobe of mice. These mice developed orthotopic tumors that were non-invasively monitored in vivo and further analyzed for tumor volume, weight, mouse survival, and immune infiltration. Further, orthotopic tumor-bearing mice were surgically castrated, leading to immediate tumor regression and subsequent recurrence, representing castration-resistant prostate cancer. Although technical skill is required to carry out this procedure, this syngeneic orthotopic model of both androgen-dependent and castration-resistant prostate cancer is of great use to all investigators in the field.

The goal of this protocol is to demonstrate the intra-prostatic injection of prostate cancer cells, with subsequent castration. Orthotopic pre-clinical models of androgen-dependent and castration-resistant prostate cancer are critical to study the disease in the context of a clinically relevant tumor microenvironment and an immunocompetent host.

アンドロゲン依存性去勢抵抗性前立腺癌の生物発光と蛍光の同所性同種同系マウスモデル

Orthotopic tumor modeling is a valuable tool for pre-clinical prostate cancer research, as it has multiple advantages over both subcutaneous and ...

Virus Delivery of CRISPR Guides to the Murine Prostate for Gene Alteration

With an increasing incidence of prostate cancer, identification of new tumor drivers or modulators is crucial. Genetically engineered mouse models (GEMM) for prostate cancer are hampered by tumor heterogeneity and its complex microevolution dynamics. Traditional prostate cancer mouse models include, amongst others, germline and conditional knockouts, transgenic expression of oncogenes, and xenograft models. Generation of de novo mutations in these models is complex, time-consuming, and costly. In addition, most of traditional models target the majority of the prostate epithelium, whereas human prostate cancer is well known to evolve as an isolated event in only a small subset of cells. Valuable models need to simulate not only prostate cancer initiation, but also progression to advanced disease.
Here we describe a method to target a few cells in the prostate epithelium by transducing cells by viral particles. The delivery of an engineered virus to the murine prostate allows alteration of gene expression in the prostate epithelia. Virus type and quantity will hereby define the number of targeted cells for gene alteration by transducing a few cells for cancer initiation and many cells for gene therapy. Through surgery-based injection in the anterior lobe, distal from the urinary track, the tumor in this model can expand without impairing the urinary function of the animal. Furthermore, by targeting only a subset of prostate epithelial cells the technique enables clonal expansion of the tumor, and therefore mimics human tumor initiation, progression, as well as invasion through the basal membrane.
This novel technique provides a powerful prostate cancer model with improved physiological relevance. Animal suffering is limited, and since no additional breeding is required, overall animal count is reduced. At the same time, analysis of new candidate genes and pathways is accelerated, which in turn is more cost efficient.

This protocol describes a newly established method for virus delivery to the murine prostate. Using either CRISPR/Cas9 technology, gene overexpression, or Cre recombinase delivery, the technique allows orthotopic alteration of gene expression and implements a novel mouse model for prostate cancer.

ウイルスの遺伝子改変マウスの前立腺に CRISPR ガイドの配信

With an increasing incidence of prostate cancer, identification of new tumor drivers or modulators is crucial. Genetically engineered mouse models (GEMM) ...

Identification, Histological Characterization, and Dissection of Mouse Prostate Lobes for In Vitro 3D Spheroid Culture Models

Genetically engineered mouse models (GEMMs) serve as effective pre-clinical models for investigating most types of human cancers, including prostate cancer (PCa). Understanding the anatomy and histology of the mouse prostate is important for the efficient use and proper characterization of such animal models. The mouse prostate has four distinct pairs of lobes, each with their own characteristics. This article demonstrates the proper method of dissection and identification of mouse prostate lobes for disease analysis. Post-dissection, the prostate cells can be further cultured in vitro for mechanistic understanding. Since mouse prostate primary cells tend to lose their normal characteristics when cultured in vitro, we outline here a method for isolating the cells and growing them as 3D spheroid cultures, which is effective for preserving the physiological characteristics of the cells. These 3D cultures can be used for analyzing cell morphology and behavior in near-physiological conditions, investigating altered levels and localizations of key proteins and pathways involved in the development and progression of a disease, and looking at responses to drug treatments.

Genetically engineered mice are useful models for investigating prostate cancer mechanisms. Here we present a protocol to identify and dissect prostate lobes from a mouse urogenital system, differentiate them based on histology, and isolate and culture the primary prostate cells in vitro as spheroids for downstream analyses.

培養 3 D 回転楕円体モデルでのマウス前立腺の葉の解剖組織学的特性及び

Genetically engineered mouse models (GEMMs) serve as effective pre-clinical models for investigating most types of human cancers, including prostate ...

MR Molecular Imaging of Prostate Cancer with a Small Molecular CLT1 Peptide Targeted Contrast Agent

Tumor extracellular matrix has abundance of cancer related proteins that can be used as biomarkers for cancer molecular imaging. In this work, we demonstrated effective MR cancer molecular imaging with a small molecular peptide targeted Gd-DOTA monoamide complex as a targeted MRI contrast agent specific to clotted plasma proteins in tumor stroma. We performed the experiment of evaluating the effectiveness of the agent for non-invasive detection of prostate tumor with MRI in a mouse orthotopic PC-3 prostate cancer model. The targeted contrast agent was effective to produce significant tumor contrast enhancement at a low dose of 0.03 mmol Gd/kg. The peptide targeted MRI contrast agent is promising for MR molecular imaging of prostate tumor.

To demonstrate MR cancer molecular imaging with a small peptide targeted MRI contrast agent specific to clotted plasma proteins in tumor stroma in a mouse prostate cancer model.

低分子CLT1ペプチド標的造影剤を用いて前立腺がんの分子イメージングのMR

Tumor extracellular  matrix has abundance of cancer related proteins that can be used as biomarkers  for cancer molecular imaging. In this work,  we ...

医学

Renal Capsule Xenografting and Subcutaneous Pellet Implantation for the Evaluation of Prostate Carcinogenesis and Benign Prostatic Hyperplasia

New therapies for two common prostate diseases, prostate cancer (PrCa) and benign prostatic hyperplasia (BPH), depend critically on experiments evaluating their hormonal regulation. Sex steroid hormones (notably androgens and estrogens) are important in PrCa and BPH; we probe their respective roles in inducing prostate growth and carcinogenesis in mice with experiments using compressed hormone pellets. Hormone and/or drug pellets are easily manufactured with a pellet press, and surgically implanted into the subcutaneous tissue of the male mouse host. We also describe a protocol for the evaluation of hormonal carcinogenesis by combining subcutaneous hormone pellet implantation with xenografting of prostate cell recombinants under the renal capsule of immunocompromised mice. Moreover, subcutaneous hormone pellet implantation, in combination with renal capsule xenografting of BPH tissue, is useful to better understand hormonal regulation of benign prostate growth, and to test new therapies targeting sex steroid hormone pathways.

We describe the manufacture of compressed hormone pellets, as well as subcutaneous surgical implantation into mice. This strategy can be combined with the growth of cell and tissue xenografts under the renal capsule of athymic nude mice to evaluate hormonal carcinogenesis and regulation of benign prostate growth.

前立腺発がんおよび良性前立腺過形成の評価のための腎皮膜異種移植および皮下ペレット移植

New therapies for two common prostate diseases,  prostate cancer (PrCa) and benign prostatic hyperplasia (BPH), depend  critically on experiments ...

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  ...

Intra-Cardiac Injection of Human Prostate Cancer Cells to Create a Bone Metastasis Xenograft Mouse Model

As the most common male malignancy, prostate cancer (PC) ranks second in mortality, primarily due to a 65%-75% bone metastasis rate. Therefore, it is essential to understand the process and related mechanisms of prostate cancer bone metastasis for developing new therapeutics. For this, an animal model of bone metastasis is an essential tool. Here, we report detailed procedures to generate a bone metastasis mouse model via intra-cardiac injection of prostate cancer cells. A bioluminescence imaging system can determine whether prostate cancer cells have been accurately injected into the heart and monitor cancer cell metastasis since it has great advantages in monitoring metastatic lesion development. This model replicates the natural development of disseminated cancer cells to form micro-metastases in the bone and imitates the pathological process of prostate cancer bone metastasis. It provides an effective tool for further exploration of the molecular mechanisms and the in vivo therapeutic effects of this disease.

Here, we present a protocol for the intra-cardiac injection of human prostate cancer cells to generate a mouse model with bone metastasis lesions.

骨転移異種移植マウスモデルを作成するためのヒト前立腺癌細胞の心臓内注射

As the most common male malignancy, prostate cancer (PC) ranks second in mortality, primarily due to a 65%-75% bone metastasis rate. Therefore, it is ...

Generation of Prostate Cancer Patient Derived Xenograft Models from Circulating Tumor Cells

Patient derived xenograft (PDX) models are gaining popularity in cancer research and are used for preclinical drug evaluation, biomarker identification, biologic studies, and personalized medicine strategies. Circulating tumor cells (CTC) play a critical role in tumor metastasis and have been isolated from patients with several tumor types. Recently, CTCs have been used to generate PDX experimental models of breast and prostate cancer. This manuscript details the method for the generation of prostate cancer PDX models from CTCs developed by our group. Advantages of this method over conventional PDX models include independence from surgical sample collection and generating experimental models at various disease stages. Density gradient centrifugation followed by red blood cell lysis and flow cytometry depletion of CD45 positive mononuclear cells is used to enrich CTCs from peripheral blood samples collected from patients with metastatic disease. The CTCs are then injected into immunocompromised mice; subsequently generated xenografts can be used for functional studies or harvested for molecular characterization. The primary limitation of this method is the negative selection method used for CTC enrichment. Despite this limitation, the generation of PDX models from CTCs provides a novel experimental model to be applied to prostate cancer research.

This manuscript details a method used to generate prostate cancer patient derived xenografts (PDXs) from circulating tumor cells (CTCs). The generation of PDX models from CTCs provides an alternative experimental model to study prostate cancer; the most commonly diagnosed tumor and a frequent cause of death from cancer in men.

前立腺癌患者の発生は、循環腫瘍細胞からの異種移植モデルを派生します

Patient derived xenograft (PDX) models are gaining popularity in cancer research and are used for preclinical drug evaluation, biomarker identification, ...

Murine Prostate Micro-dissection and Surgical Castration

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has been used as a surrogate for discoveries in human prostate development and disease. Prostate micro-dissection allows consistent study of lobe-specific prostate anatomy, histology, and cellular characteristics in the absence of contamination of other tissues. Testosterone affects prostate development and disease. Androgen deprivation therapy is a common treatment for prostate cancer patients, but many prostate tumors become castration-resistant. Surgical castration of mouse models allows for the study of castration resistance and other facets of hormonal biology on the prostate. This procedure can be coupled with testosterone reintroduction, or hormonal regeneration of the prostate, a powerful method to study stem cell lineages in the prostate. Together, prostate micro-dissection and surgical castration opens up a multitude of opportunities for robust and consistent research of prostate development and disease. This manuscript describes the protocols for prostate micro-dissection and surgical castration in the laboratory mouse.

This manuscript describes the protocols for prostate micro-dissection and surgical castration in the laboratory mouse. We also depict representative results produced by these protocols. Finally, we discuss the advantages and utilization of these protocols.

マウス前立腺マイクロ解剖と外科的去勢

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has ...