国立衛生研究所（KLへCA133085）と米国癌学会からの補助金（KLにRSG - 09 - 209 - 01 - TBG）によってサポートされています。

1。実験的転移モデルマウス<ol><li>腫瘍細胞の注射の前日に、シードone T75フラスコ上で1 × 10 7〜急成長中の腫瘍細胞を得るために、10％血清を含むRPMI培地10mLに細胞をCMS4 - MET。 37℃で一晩インキュベート℃、 </li><li>注射の日に、培地を除去し、PBSで細胞を1回洗い流した後、37℃で0.05％トリプシン- EDTAで腫瘍細胞を収穫℃で5分間。 10％血清を含むRPMI培地10mlで反応を停止します。コニカルチューブに細胞を移す。 </li><li>室温で3分間ソーバルレジェンドRT遠心機で1300 rpmで細胞をスピンダウン。上清を取り除く。洗浄に10mLの新鮮な1X HBSSで細胞を再懸濁し、再度スピンダウンと同じように再懸濁します。 </li><li>血球計算板上で腫瘍細胞を数える。それぞれの注入に必要な細胞を100μLの全体積に再懸濁しているように、HBSSで細胞を希釈する。 </li><li>尾静脈を拡張させるために温水に浸漬ビーカーに6〜8週齢のBALB / cマウスを温める。ベンチ上で尾静脈の現像抑制剤の上にマウスを置きます。外側尾静脈内に腫瘍細胞を100μLを注入するためにマイクロシリンジを使用してください。感染を避けるために、無菌操作を使用してください。出血が停止するまで注入した後、注射部位にわずかな圧力を適用する。 </li><li>ケージにマウスを返し、そして腫瘍が目的の段階に成長することができます。 </li></ol> 2。細胞傷害性Tリンパ球（CTL）の養子移入免疫療法<ol><li>移転の日に、精製されたT -リンパ球細胞傷害性、またはCTLは、テキストで説明したように調製を再懸濁するために上下にピペッティングし。つのプレートから15 mLコニカルチューブに細胞のすべてを譲渡する、ボリュームがチューブの2 / 3以上を超えてさせることではない。 </li><li>コニカルチューブに滅菌パスツールピペットを挿入し、総容量は14 mLのに近づくまで、セルの下に、リンパ球分離媒体、またはLSMを置く。 </li><li>層を乱さないように注意してください。室温で20分間2000rpmで遠心する。スピンした後ローターを減速させるブレーキを使用しないでください。 </li><li>新しい15 mLコニカルチューブにCTLを転送する。洗浄に約10mlの体積にHBSSを追加。 </li><li>細胞を数える。室温で5分間1300 rpmでスピンダウン。 100μLに注入あたりの​​総容積を保ち、注射のために必要な細胞密度にHBSSに再懸濁します。 </li><li>担癌マウスにCTLを注入する前のこのビデオで見たのと同じテールのインジェクションの手法を使用してください。 </li><li>ケージにマウスを返す、とCTLsは腫瘍と対話することができます。マウスは、通常14-21日CTL治療後の分析のために犠牲にされています。 </li></ol> 3。肺転移の可視化<ol><li>肺転移を可視化するために、発泡スチロールのボードに、その裏に犠牲にマウスを置きます。気管に遮るもののないアクセスを確保するために足を固定。 70％エタノールで腹側を吹き付けます。 </li><li>半ば腹部から始まる、正中線に沿って、胸郭を介して、とまで唾液腺に向かってカットするはさみを使用してください。気管を公開。気管の周囲の組織を削除するには、ピンセットを使用してください。 </li><li>気管の下に200μLピペットの先端を通します。片手で先端を保持する、静かに体から気管を持ち上げて。 </li><li>プラットフォームを180 °回転させる。気管から肺にインドのインクを注入するために50mLのシリンジを使用してください。あなたが強い抵抗を感じるまで、完全にインクで肺を膨らませる。 </li><li>気管をカットするはさみを使用してください。肺の下にピンセットをスライドさせ、それらをマウスから持ち上げます。水の1Lビーカーに簡単に肺をすすいでください。 </li><li>化学煙フードでは、フェケテの液5 mLを含有するガラスシンチレーションバイアルに肺を転送する。腫瘍組織は、数分後に黒の肺に白い結節として現れるだろう。腫瘍は、現在カウントし、無期限にフェケテのソリューションに格納することができます。 </li></ol> 4。視覚による肺の代表的な転移<ol><li>ここでは、乳癌細胞株4T1を注入したマウスからの肺には腫瘍を示す白い斑点を示す。 IRF8ドミナントネガティブ変異体K79Eでトランスフェクトした4T1細胞を注入したマウスは、腫瘍細胞の拡張された転移能を示す。 </li><li>これらのイメージは、CTLの養子移入免疫療法の有効性の組織学的分析を示しています。生理食塩水を注射した担癌マウスでは、黄色の矢印（）で示される、6日後に複数の腫瘍を認めた。腫瘍特異的T細胞を注射したマウスは、その一方で、腫瘍（B）の減少を示した。 </li><li>同じ実験では、インドのインクの治療は、CTL養子免疫伝達の有効性を測定する簡単な方法を提供します。ここでは、担癌肺の白い腫瘍結節は、明らかに成功したCTLの養子移入を受けた肺からそれらを区別し、結節を数える度を可能に組織学と定量の可能性ではない。 </li></ol> 5。代表的な結果<img src="/files/ftp_upload/2077/2077fig1.jpg" alt="図1" /> 図1実験的な腫瘍転移およびCTL適応転送免疫療法のマウスモデルの実験スキーム。赤い点は、腫瘍細胞と緑のドットはCTLを示して示している。 <img src="/files/ftp_upload/2077/2077fig2.jpg" alt="図2" /> 図2。IRF8機能の破壊は、腫瘍細胞の転移能を増強した。マウス乳癌細胞株4T1は安定してベクトル（4T1.Vector）またはIRF8ドミナントネガティブ変異体K79E（4T1.IRF8K79E）（15,16）を発現ベクターでトランスフェクトした。腫瘍細胞は、マウスの外側尾静脈に静脈注射された。腫瘍のある肺は腫瘍結節を視覚化するインドのインクで膨らませていた。腫瘍結節は、簡単に黒の肺組織の背景に白い斑点として見られている。 <img src="/files/ftp_upload/2077/2077fig3.jpg" alt="図3" /> 図3。CTLの養子移入免疫療法の有効性の組織学的分析。マウス肉腫細胞株CMS4 - Metはマウス側面尾静脈に静脈注射した。三日後、生理食塩水（）または腫瘍特異的T細胞は、（B）担癌マウスに注入した。肺は6日、従来のH＆E組織染色により、CTL治療後に分析した。腫瘍結節は、矢印で示されている。 <img src="/files/ftp_upload/2077/2077fig4.jpg" alt="図4" /> 図4。水墨インフレによる腫瘍結節の可視化。マウス肉腫細胞株CMS4 - Metはマウス側面尾静脈に静脈注射した。三日後、生理食塩水（コントロール）または腫瘍特異的T細胞は、（+ CTL）担癌マウスに注入した。肺は14日CTL治療後に分析した。腫瘍結節ている白い点は、、CTL治療の有効性を簡単に定量化が可能になります。

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マウスは、国立癌研究所（Friderick、MD）から購入し、ジョージア州の動物施設の医科大学に収容された。実験と医療/福祉は連邦政府の規制との契約とグルジアIACUC委員会の医科大学によって承認されたプロトコルでいた。

ソリューション： インドのインク液（17）： <ol><li> 250mlのフラスコに蒸留水150mlを注ぐ。 </li><li> 4滴を蒸留水に水酸化アンモニウムを追加。 </li><li>アンモニアと水の混合物に30ミリリットルインドのインク株式（すなわちサンフォードブラックマジック防水ドローイングインク4465アイテム44011）を追加します。 </li><li> 200mlの量を蒸留水で仕上げる。解決策は、注射の準備ができています。 </li></ol>フェケテのソリューション（17）： フェケテのソリューションは、正常組織の背景を黒から白の腫瘍結節を区別するために漂白剤墨汁-水増し担癌肺に使用されます。 <ol><li> 1Lガラス瓶に350ミリリットルの95％EtOHを追加します。 </li><li> 150ミリリットルの蒸留水を追加します。 </li><li> 50ミリリットルホルムアルデヒドを追加</li><li> 25ミリリットルの氷酸性酸を追加</li></ol>

experimental metastasis and ctl adoptive transfer immunotherapy mouse model

実験的転移のマウスモデルは、生理学的に関連はまだシンプルと転移モデルです。循環の生存率、遠位臓器の血管外漏出と植民地化：腫瘍細胞は腫瘍細胞自然転移の最後のステップに似て、それによって、マウスの尾静脈に（IV）静脈内に注射し、肺に植民地化されています。遠位臓器で確立された転移性腫瘍：治療の目標はしばしば転移のエンドポイントであるので、治療的観点から、実験的転移モデルは最も単純で理想的なモデルです。このモデルでは、腫瘍細胞は、マウスの尾静脈に静脈注射し、肺に植民地化と拡張されます。腫瘍特異的CTLは、その後、転移、担癌マウスに静脈注射されています。肺転移の数と大きさが注入される腫瘍細胞の数と腫瘍の成長の時間によって制御することができます。したがって、転移の様々な段階では、広範な転移への最小限の転移から、モデル化することができます。肺転移は、このように簡単に目視観察および定量を可能に、インクでインフレによって分析されています。

Watch this Scientific Journal Video about Experimental Metastasis and CTL Adoptive Transfer Immunotherapy Mouse Model at JoVE.com

Experimental Metastasis and CTL Adoptive Transfer Immunotherapy Mouse Model

腫瘍細胞- T細胞相互作用の分析のための実験的肺転移とCTL免疫療法のマウスモデル<em&gt; in vivoで</em&gt;。

実験的転移とCTLの養子移入免疫療法のマウスモデル

Experimental metastasis mouse model is a simple and yet physiologically relevant metastasis model. The tumor cells are injected intravenously (i.v) into ...

experimental-metastasis-ctl-adoptive-transfer-immunotherapy-mouse

Nanyang Technological University

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Immunology and Infection

19.3K ビュー.  Medical College of Georgia. 腫瘍細胞- T細胞相互作用の分析のための実験的肺転移とCTL免疫療法のマウスモデル in vivoで。

ビデオ: Experimental Metastasis and CTL Adoptive Transfer Immunotherapy Mouse Model

Advanced Animal Model of Colorectal Metastasis in Liver: Imaging Techniques and Properties of Metastatic Clones

Patients with a limited number of hepatic metastases and slow rates of progression can be successfully treated with local treatment approaches1,2. However, little is known about the heterogeneity of liver metastases, and animal models capable of evaluating the development of individual metastatic colonies are needed. Here, we present an advanced model of hepatic metastases that provides the ability to quantitatively visualize the development of individual tumor clones in the liver and estimate their growth kinetics and colonization efficiency. We generated a panel of monoclonal derivatives of HCT116 human colorectal cancer cells stably labeled with luciferase and tdTomato and possessing different growth properties. With a splenic injection followed by a splenectomy, the majority of these clones are able to generate hepatic metastases, but with different frequencies of colonization and varying growth rates. Using the In Vivo&#160;Imaging System (IVIS), it is possible to visualize and quantify metastasis development with in vivo luminescent and ex vivo fluorescent imaging. In addition, Diffuse Luminescent Imaging Tomography (DLIT) provides a 3D distribution of liver metastases in vivo. Ex vivo fluorescent imaging of harvested livers provides quantitative measurements of individual hepatic metastatic colonies, allowing for the evaluation of the frequency of liver colonization and the growth kinetics of metastases. Since the model is similar to clinically observed liver metastases, it can serve as a modality for detecting genes associated with liver metastasis and for testing potential ablative or adjuvant treatments for liver metastatic disease.

The ability of metastatic clones to colonize distant sites depends on their proliferation capacity and/or their ability to survive in the host microenvironment without significant proliferation. Here, we present an animal model that allows quantitative visualization of both types of liver colonization by metastatic clones.

肝臓における大腸転移の高度な動物モデル：イメージング技術および転移性クローンのプロパティ

Patients with a limited number of hepatic metastases and slow rates of progression can be successfully treated with local treatment approaches1,2. ...

がん研究

A Syngeneic Mouse Model of Metastatic Renal Cell Carcinoma for Quantitative and Longitudinal Assessment of Preclinical Therapies

Renal cell carcinoma (RCC) affects &#62; 60,000 people in the United States annually, and ~ 30% of RCC patients have multiple metastases at the time of diagnosis. Metastatic RCC (mRCC) is incurable, with a median survival time of only 18 months. Immune-based interventions (e.g., interferon (IFN) and interleukin (IL)-2) induce durable responses in a fraction of mRCC patients, and multikinase inhibitors (e.g., sunitinib or sorafenib) or anti-VEGF receptor monoclonal antibodies (mAb) are largely palliative, as complete remissions are rare. Such shortcomings in current therapies for mRCC patients provide the rationale for the development of novel treatment protocols. A key component in the preclinical testing of new therapies for mRCC is a suitable animal model. Beneficial features that recapitulate the human condition include a primary renal tumor, renal tumor metastases, and an intact immune system to investigate any therapy-driven immune effector responses and the formation of tumor-induced immunosuppressive factors. This report describes an orthotopic mRCC mouse model that has all of these features. We describe an intrarenal implantation technique using the mouse renal adenocarcinoma cell line Renca, followed by the assessment of tumor growth in the kidney (primary site) and lungs (metastatic site).

Implementation of an orthotopic model of renal cell carcinoma in immunocompetent mice affords the investigator a clinically-relevant system defined by the presence of a primary renal tumor and lung metastases in the same animal. This system can be used to preclinically test a variety of treatments in vivo.

定量的および前臨床治療の縦評価のための転移性腎細胞癌の同系マウスモデル

Renal cell carcinoma (RCC) affects &#62; 60,000 people in the United States annually, and ~ 30% of RCC patients have multiple metastases at the time of ...

A Murine Ommaya Xenograft Model to Study Direct-Targeted Therapy of Leptomeningeal Disease

Leptomeningeal disease (LMD) is an uncommon type of central nervous system (CNS) metastasis to the cerebral spinal fluid (CSF). The most common cancers that cause LMD are breast and lung cancers and melanoma. Patients diagnosed with LMD have a very poor prognosis and generally survive for only a few weeks or months. One possible reason for the lack of efficacy of systemic therapy against LMD is the failure to achieve therapeutically effective concentrations of drug in the CSF because of an intact and relatively impermeable blood-brain barrier (BBB) or blood-CSF barrier across the choroid plexus. Therefore, directly administering drugs intrathecally or intraventricularly may overcome these barriers. This group has developed a model that allows for the effective delivery of therapeutics (i.e., drugs, antibodies, and cellular therapies) chronically and the repeated sampling of CSF to determine drug concentrations and target modulation in the CSF (when the tumor microenvironment is targeted in mice). The model is the murine equivalent of a magnetic resonance imaging-compatible Ommaya reservoir, which is used clinically. This model, which is affixed to the skull, has been designated as the &#34;Murine Ommaya.&#34; As a therapeutic proof of concept, human epidermal growth factor receptor 2 antibodies (clone 7.16.4) were delivered into the CSF via the Murine Ommaya to treat mice with LMD from human epidermal growth factor receptor 2-positive breast cancer. The Murine Ommaya increases the efficiency of drug delivery using a miniature access port and prevents the wastage of excess drug; it does not interfere with CSF sampling for molecular and immunological studies. The Murine Ommaya is useful for testing novel therapeutics in experimental models of LMD.

Here, we describe a murine xenograft model that functionally resembles an Ommaya reservoir in patients. We developed the Murine Ommaya to study novel therapeutics for the universally fatal leptomeningeal disease.

レプトメニンゲレス病の直接標的療法を研究するマウス・オンマヤ異種移植片モデル

Leptomeningeal disease (LMD) is an uncommon type of central nervous system (CNS) metastasis to the cerebral spinal fluid (CSF). The most common cancers ...

Tumor Transplantation for Assessing the Dynamics of Tumor-Infiltrating CD8+ T Cells in Mice

T cell-mediated immunity plays a crucial role in immune responses against tumors, with cytotoxic T lymphocytes (CTLs) playing the leading role in eradicating cancerous cells. However, the origins and replenishment of tumor antigen-specific CD8+ T cells within the tumor microenvironment (TME) remain obscure. This protocol employs the B16F10-OVA melanoma cell line, which stably expresses the surrogate neoantigen, ovalbumin (OVA), and TCR transgenic OT-I mice, in which over 90% of CD8+ T cells specifically recognize the OVA-derived peptide OVA257-264 (SIINFEKL) bound to the class I major histocompatibility complex (MHC) molecule H2-Kb. These features enable the study of antigen-specific T cell responses during tumorigenesis.
Combining this model with tumor transplantation surgery, tumor tissues from donors were transplanted into tumor-matched syngeneic recipient mice to precisely trace the influx of recipient-derived immune cells into transplanted donor tissues, allowing the analysis of the immune responses of tumor-inherent and periphery-originated antigen-specific CD8+ T cells. A dynamic transition was found to occur between these two populations. Collectively, this experimental design has provided another approach to precisely investigate the immune responses of CD8+ T cells in TME, which will shed new light on tumor immunology.

Tumor Transplantation for Assessing the Dynamics of Tumor-Infiltrating CD8+ T Cells in Mice

Here, we present a tumor transplantation protocol for the characterization of tumor-inherent and periphery-derived tumor-infiltrated lymphocytes in a mouse tumor model. Specific tracing of the influx of recipient-derived immune cells with flow cytometry reveals the dynamics of the phenotypic and functional changes of these cells during antitumor immune responses.

マウスにおける腫瘍浸潤CD8+ T細胞の動態を評価するための腫瘍移植

T cell-mediated immunity plays a crucial role in immune responses against tumors, with cytotoxic T lymphocytes (CTLs) playing the leading role in ...

Induction of Invasive Transitional Cell Bladder Carcinoma in Immune Intact Human MUC1 Transgenic Mice:  A Model for Immunotherapy Development

A preclinical model of invasive bladder cancer was developed in human mucin 1 (MUC1) transgenic (MUC1.Tg) mice for the purpose of evaluating immunotherapy and/or cytotoxic chemotherapy. To induce bladder cancer, C57BL/6 mice (MUC1.Tg and wild type) were treated orally with the carcinogen N-butyl-N-(4-hydroxybutyl)nitrosamine (OH-BBN) at 3.0 mg/day, 5 days/week for 12 weeks. To assess the effects of OH-BBN on serum cytokine profile during tumor development, whole blood was collected via submandibular bleeds prior to treatment and every four weeks. In addition, a MUC1-targeted peptide vaccine and placebo were administered to groups of mice weekly for eight weeks. Multiplex fluorometric microbead immunoanalyses of serum cytokines during tumor development and following vaccination were performed. At termination, interferon gamma (IFN-&#947;)/interleukin-4 (IL-4) ELISpot analysis for MUC1 specific T-cell immune response and histopathological evaluations of tumor type and grade were performed. The results showed that: (1) the incidence of bladder cancer in both MUC1.Tg and wild type mice was 67%; (2) transitional cell carcinomas (TCC) developed at a 2:1 ratio compared to squamous cell carcinomas (SCC); (3) inflammatory cytokines increased with time during tumor development; and (4) administration of the peptide vaccine induces a Th1-polarized serum cytokine profile and a MUC1 specific T-cell response. All tumors in MUC1.Tg mice were positive for MUC1 expression, and half of all tumors in MUC1.Tg and wild type mice were invasive. In conclusion, using a team approach through the coordination of the efforts of pharmacologists, immunologists, pathologists and molecular biologists, we have developed an immune intact transgenic mouse model of bladder cancer that expresses hMUC1.

An N-butyl-N-(4-hydroxybutyl)nitrosamine-induced bladder cancer model was developed in human mucin 1 (MUC1) transgenic mice for the purpose of testing MUC1-directed immunotherapy. After administering a MUC1-targeted peptide vaccine, a cytotoxic T lymphocyte response to MUC1 was confirmed by measuring serum cytokine levels and T-cell specific activity.

免疫無傷のヒトMUC1トランスジェニックマウスにおける侵襲移行細胞膀胱癌の誘導：免疫療法の開発のためのモデル

A preclinical model of invasive bladder cancer was developed in human mucin 1 (MUC1) transgenic (MUC1.Tg) mice for the purpose of evaluating immunotherapy ...

医学

A Robust Discovery Platform for the Identification of Novel Mediators of Melanoma Metastasis

Metastasis is a complex process, requiring cells to overcome barriers that are only incompletely modeled by in vitro assays. A systematic workflow was established using robust, reproducible in vivo models and standardized methods to identify novel players in melanoma metastasis. This approach allows for data inference at specific experimental stages to precisely characterize a gene's role in metastasis. Models are established by introducing genetically modified melanoma cells via intracardiac, intradermal, or subcutaneous injections into mice, followed by monitoring with serial in vivo imaging. Once preestablished endpoints are reached, primary tumors and/or metastases-bearing organs are harvested and processed for various analyses. Tumor cells can be sorted and subjected to any of several 'omics' platforms, including single-cell RNA sequencing. Organs undergo imaging and immunohistopathological analyses to quantify the overall burden of metastases and map their specific anatomic location. This optimized pipeline, including standardized protocols for engraftment, monitoring, tissue harvesting, processing, and analysis, can be adopted for patient-derived, short-term cultures and established human and murine cell lines of various solid cancer types.

This article describes a workflow of techniques employed for testing novel candidate mediators of melanoma metastasis and their mechanism(s) of action.

メラノーマ転移の新規メディエーターの同定のための堅牢な発見プラットフォーム

Metastasis is a complex process, requiring cells to overcome barriers that are only incompletely modeled by in vitro assays. A systematic workflow was ...

Orthotopic Implantation of Patient-Derived Cancer Cells in Mice Recapitulates Advanced Colorectal Cancer

Over the last decade, more sophisticated preclinical colorectal cancer (CRC) models have been established using patient-derived cancer cells and 3D tumoroids. Since patient derived tumor organoids can retain the characteristics of the original tumor, these reliable preclinical models enable cancer drug screening and the study of drug resistance mechanisms. However, CRC related death in patients is mostly associated with the presence of metastatic disease. It is therefore essential to evaluate the efficacy of anti-cancer therapies in relevant in vivo models that truly recapitulate the key molecular features of human cancer metastasis. We have established an orthotopic model based on the injection of CRC patient-derived cancer cells directly into the cecum wall of mice. These tumor cells develop primary tumors in the cecum that metastasize to the liver and lungs, which is frequently observed in patients with advanced CRC. This CRC mouse model can be used to evaluate drug responses monitored by microcomputed tomography (&#181;CT), a clinically relevant small-scale imaging method that can easily identify primary tumors or metastases in patients. Here, we describe the surgical procedure and the required methodology to implant patient-derived cancer cells in the cecum wall of immunodeficient mice.

This protocol describes the orthotopic implantation of patient-derived cancer cells in the cecum wall of immunodeficient mice. The model recapitulates advanced colorectal cancer metastatic disease and allows for the evaluation of new therapeutic drugs in a clinically relevant scenario of lung and liver metastases.

進行性大腸癌を再現するマウスへの患者由来癌細胞の同所性移植

Over the last decade, more sophisticated preclinical colorectal cancer (CRC) models have been established using patient-derived cancer cells and 3D ...

Experimental Melanoma Immunotherapy Model Using Tumor Vaccination with a Hematopoietic Cytokine

Fms-like tyrosine kinase 3 ligand (Flt3L) is a hematopoietic cytokine that promotes the survival and differentiation of dendritic cells (DCs). It has been used in tumor vaccines to activate innate immunity and enhance antitumor responses. This protocol demonstrates a therapeutic model using cell-based tumor vaccine consisting of Flt3L-expressing B16-F10 melanoma cells along with phenotypic and functional analysis of immune cells in the tumor microenvironment (TME). Procedures for cultured tumor cell preparation, tumor implantation, cell irradiation, tumor size measurement, intratumoral immune cell isolation, and flow cytometry analysis are described. The overall goal of this protocol is to provide a preclinical solid tumor immunotherapy model, and a research platform to study the relationship between tumor cells and infiltrating immune cells. The immunotherapy protocol described here can be combined with other therapeutic modalities, such as immune checkpoint blockade (anti-CTLA-4, anti-PD-1, anti-PD-L1 antibodies) or chemotherapy in order to improve the cancer therapeutic effect of melanoma.

The protocol presents a cancer immunotherapy model using cell-based tumor vaccination with Flt3L-expressing B16-F10 melanoma. This protocol demonstrates the procedures, including preparation of cultured tumor cells, tumor implantation, cell irradiation, measurement of tumor growth, isolation of intratumoral immune cells, and flow cytometry analysis.

造血サイトカインによる腫瘍ワクチン接種を用いた実験的黒色腫免疫療法モデル

Fms-like tyrosine kinase 3 ligand (Flt3L) is a hematopoietic cytokine that promotes the survival and differentiation of dendritic cells (DCs). It has been ...

A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of cancer metastasis are still unclear. Animal models are essential for elucidating the mechanisms and for evaluating novel strategies for the treatment of metastatic cancers. Here, an in-depth description of a &#8220;patient-like&#8221; orthotopic syngeneic mouse model for exploring the mechanisms of metastasis of solid organ tumors is provided. The survival surgical implantation of BNL 1ME A.7R.1 mouse hepatocellular carcinoma cells directly into the liver (the organ of origin) of the inbred wild-type immune competent laboratory mouse strain, BALB/c is described. The success and reproducibility of this methodology recommends it for widespread use in elucidating the biological mechanisms of solid organ cancer metastasis.

The metastatic spread of cancer is the major cause of cancer-related deaths. We provide an in-depth description of our survival surgery methodology for establishing a &#8220;patient-like&#8221; orthotopic syngeneic mouse model system for studying the mechanisms of metastasis in solid organ tumors.

肝細胞癌転移の「患者様」同所同系マウスモデル

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of ...

<meta charset="utf8"></head><body>膵臓がんモデルにおける腫瘍浸潤リンパ球を用いた養子縁組療法

Flow Cytometry-Based Isolation and Therapeutic Evaluation of Tumor-Infiltrating Lymphocytes in a Mouse Model of Pancreatic Cancer

Pancreatic cancer is an aggressive malignancy with a dismal prognosis and limited therapeutic options. Adoptive cell therapy, which involves isolating and activating a patient&#39;s own immune cells, such as tumor-infiltrating lymphocytes (TILs), before re-infusing them, represents a promising experimental approach. However, techniques for adoptive cell transfer in preclinical pancreatic cancer models are not well established. Here, we describe a detailed protocol for adoptive cell therapy using TILs from a syngeneic pancreatic cancer mouse model. The procedure involves implanting live or irradiated mouse pancreatic cancer cells in fluorescence-labeled reporter mice to initiate immune cell influx, then isolating lymphocytes from primary tumors via flow cytometry sorting and/or activating and expanding tumor-reactive T cells ex vivo, and adoptively transferring these activated T cells intraperitoneally into tumor-bearing mice, followed by interleukin-2 administration.&#160;Bioluminescent tumor imaging allows for longitudinal monitoring of orthotopic tumor growth and response to therapy, especially evaluating the tumor-specific cytotoxic effects. This approach recapitulates the logistics involved in developing adoptive cell transfer therapies for pancreatic cancer patients. The results demonstrate enhanced antitumor efficacy of adoptively transferred tumor-reactive T cells compared to irrelevant lymphocyte controls. This versatile methodology enables the in vivo study of adoptive immunotherapy in pancreatic cancer as well as the optimization of cell processing parameters and combination treatment regimens.

<meta charset="utf8"></head><body>膵臓癌マウスモデルにおける腫瘍浸潤リンパ球のフローサイトメトリーに基づく単離と治療的評価

We describe a method to utilize tumor-infiltrating lymphocytes (TILs) from mice through flow cytometry for adoptive cell transfer. This protocol aims to verify the specific cytotoxicity of TILs against tumors in a syngeneic pancreatic cancer mouse model, providing insights into the development of adoptive cell therapies for pancreatic cancer.

膵臓癌マウスモデルにおける腫瘍浸潤リンパ球のフローサイトメトリーに基づく単離と治療的評価

Pancreatic cancer is an aggressive malignancy with a dismal prognosis and limited therapeutic options. Adoptive cell therapy, which involves isolating and ...