Name: intracranial implantation with subsequent 3d in vivo bioluminescent imaging of murine gliomas
Uploaded: 2011-11-06T14:00:00.000+00:00
Duration: 9 min 46 s
Description: Watch this Scientific Journal Video about Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas at JoVE.com

我々は、GL261 - lucの細胞の調製に関する有用な提案のためのレンチウイルスのシステムだけでなく、Mahilラオのためのプラスミドの寛大な贈り物のために博士ジョシュアB.ルービンに感謝します。 我々は、脳腫瘍研究（SSBTR）、バロー神経財団とその寛大な支援のためのウォレス財団を支援する学生に感謝する。 動物実験は、セントジョセフ病院と医療センターの動物実験使用の委員会が定めるガイドラインおよび規則に従って行った。

<ol>
	<li>Candolfi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracranial+glioblastoma+models+in+preclinical+neuro-oncology:+neuropathological+characterization+and+tumor+progression.">Intracranial glioblastoma models in preclinical neuro-oncology: neuropathological characterization and tumor progression.</a> J Neurooncol. 85, 133-148 (2007).</li><li>Stafford, P., Abdelwahab, M. G., do, K. i. m., Preul, Y., Rho, M. C., M, J., Scheck, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+ketogenic+diet+reverses+gene+expression+patterns+and+redudes+reactive+oxygen+species+levels+when+used+as+an+adjuvant+therapy+for+glioma.">The ketogenic diet reverses gene expression patterns and redudes reactive oxygen species levels when used as an adjuvant therapy for glioma.</a> Nutr Metab. 7, (2010).</li><li>Caliper Life Sciences, Inc. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Living Image Software Version 4.0. VivoVision Systems. , (2010).</li><li>Paxinos, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Mouse Brain in Stereotaxic Coordinates. , (2001).</li><li>Jouanneau, E., Poujol, D., Gulia, S., Le, M. I., Blay, J. Y., Belin, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dendritic+cells+are+essential+for+priming+but+inefficient+for+boosting+antitumour+immune+response+in+an+orthotopic+murine+glioma+model.">Dendritic cells are essential for priming but inefficient for boosting antitumour immune response in an orthotopic murine glioma model.</a> Cancer Immunol Immunother. 55, 254-267 (2006).</li></ol>

この記事へのフリーアクセスは、キャリパーライフサイエンスが主催している。

細胞の播種は、0.4mmのポケットを作成した後、脳の表面から2.6 mmの深さで注入される。針の適切な配置と深さを確保するためにX線は、C -アームまたは類似のX線画像の激化デバイスを使用して取得できますが、これはオプションです。動物が適切に鎮静されていない場合、手術の合併症は、動物の細胞の注入の間に移動できる点で、発生する場合があります。これは、針のトラックからの細胞の混合または出血の漏洩を引き起こす可能性があります。セルのリーク電流は、腫瘍細胞の異所性増殖を引き起こす。またburrholeはプロトコル4で説明されている2.3ミリメートルの内側行われた場合に行うことができます心室ではない穿刺することが重要です。針と細胞拡散の適切な配置は、2μlのメチレンブルー色素でマウスを注入し、注入された色素の場所を確認するために脳組織を解剖することによって試験した。 このプロトコルでは、in vivoイメージングシステムとt で IVISスペクトラムを使用している彼はこの楽器で使用するために設計されたイメージのソフトウェア（V 4.0）リビング。 （キャリパーライフサイエンス）。 in vivoイメージングシステムと画像解析ツールであらゆる匹敵するが、同様の結果を得るために使用することができます。これらのシステムは、実験的な頭蓋内腫瘍の成長をフォローするために従来の磁気共鳴画像（MRI）に比べていくつかの利点を提供します。最も明白な2楽器の相対的なコストである-動物MRI装置は、はるかに高価であり、一般的に熟練したMRI技術者のサービスを必要とする。ここに記載されたものとin vivoイメージングでは 、エンドユーザが行うことができます。 MRIデータの定量には時間がかかり、やや不正確である間の生物発光データは、定量的である。また、MRI画像は腫瘍細胞に加えて、浮腫や炎症を表示し、それが治療効果から腫瘍を分離することが困難な場合があります。これらの理由から成長する腫瘍の正確な体積測定値を得ることが困難な場合があります。生物発光は、ATPが必要ですしたがって、唯一の生きている腫瘍細胞は、腫瘍の大きさのデータに貢献する。マシンが使用可能であればそれにもかかわらず、MRIにはいくつかの利点があります。細胞は、MRIで可視化するために生物発光マーカーで標識する必要はありません。周囲の腫瘍浮腫を視覚化する能力は、いくつかの実験的なプロトコルのための利点があります。両方の技術の強さは、いずれかを使用すると、他の使用を排除しない、両方のテクノロジが利用可能になったとき、そのデータは腫瘍の成長だけでなく、同​​じ動物からペリ腫瘍浮腫や炎症の存在を得ることができることです。研究者に。

<table border="1"><tbody><tr><td> 試薬や供給の名前 </td><td> 会社 </td><td> カタログ番号 </td><td> コメント </td></tr><tr><td> GL261 - luc2バイオウェアウルトラ</td><td>キャリパーライフサイエンス</td><td> GL261 - luc2 </td><td></td></tr><tr><td>ダルベッコ改変イーグル培地（DMEM） </td><td>インビトロジェン</td><td> 10313039 </td><td></td></tr><tr><td>ジェネティシン（G418） </td><td> GIBCO（インビトロジェン） </td><td> 11811-023 </td><td></td></tr><tr><td>ウシ胎児血清（FCS） </td><td>インビトロジェン</td><td> 26140079 </td><td></td></tr><tr><td> Phospateは緩衝生理食塩水（PBS） </td><td>インビトロジェン</td><td> 70011044 </td><td></td></tr><tr><td> C57BL/6-cBrd/cBrd/Crマウス</td><td> NCI -フレデリック</td><td></td><td></td></tr><tr><td>アクワの涙の潤滑眼軟膏</td><td> Akorn株式会社</td><td> 17478-062-35 </td><td></td></tr><tr><td> Ketaset（塩酸ケタミン） </td><td>ワイエス</td><td> 11570775 </td><td></td></tr><tr><td> Sedazine（キシラジン塩酸塩） </td><td>ワイエス</td><td> 10031894 </td><td></td></tr><tr><td>小動物の定位音源</td><td> Kopfは楽器</td><td> 900 </td><td></td></tr><tr><td> SYS - Micro4コントローラとUltraMicroPump </td><td>世界の精密機器</td><td> UMP3 - 1 </td><td>利用できない場合は、手動で注入することが可能です</td></tr><tr><td> 26ゲージと10μlのシリンジは、針の面取り</td><td>世界の精密機器</td><td> SGE010RNS </td><td></td></tr><tr><td> Adison鉗子</td><td>世界の精密機器</td><td> 500092 </td><td></td></tr><tr><td>ペンフィールド解剖</td><td>タラ漁船</td><td> 65から1015 </td><td></td></tr><tr><td> 16グラム1 ½精密グライドニードル</td><td> Beckton、ディッキンソン社（BD） </td><td> 305198 </td><td></td></tr><tr><td>外科ブレードハンドル</td><td> BD </td><td> 371030 </td><td></td></tr><tr><td>サイズ15ブレード</td><td> BD </td><td> 371315 </td><td></td></tr><tr><td> 4から0 Vicryl縫合糸</td><td>エチコン</td><td> VCP496G </td><td></td></tr><tr><td>骨ろう</td><td> MEDLINE </td><td> DYNJBW25 </td><td></td></tr><tr><td> Povidine -ヨウ素綿棒スティック</td><td> MEDLINE </td><td> MD93901 </td><td></td></tr><tr><td> D -ルシフェリンカリウム塩</td><td>キャリパーライフサイエンス</td><td> 122796 </td><td></td></tr><tr><td> Forane（イソフルラン） </td><td>バクスター</td><td> 1001936060 </td><td></td></tr><tr><td> OPMI Pentero顕微鏡</td><td>カールツァイス株式会社</td><td></td><td>どんな手術用顕微鏡で十分です</td></tr><tr><td>オプションの麻酔システムとXenogen IVISスペクトラム</td><td>キャリパーライフサイエンス</td><td></td><td></td></tr></tbody></table>

intracranial implantation with subsequent 3d in vivo bioluminescent imaging of murine gliomas

マウス神経膠腫261（GL261）はin vivoでのモデル系として認識されているヒトの多形性膠芽腫（GBM）の機能の多くは反復する。細胞株は、もともとC57BL / 6同系マウスの系統1に3 -メチル- cholantreneの頭蓋内注射によって誘発されたので、免疫学的に有能なC57BL / 6マウスを使用することができます。我々はGL261を使用していますが、次のプロトコルは、任意の頭蓋内のマウス腫瘍モデルの移植とモニタリングに使用することができます。 GL261細胞を安定的に発現するホタルルシフェラーゼ（GL261 - LUC）に設計された。我々はまた、CMVプロモーターから発現luc2遺伝子の安定なトランスフェクションにより、明るくGL261 - luc2細胞株を作成した。国立癌研究所、フレデリック、MDからC57BL/6-cBrd/cBrd/Crマウス（C57BL / 6のアルビノ変異体）は黒の皮と毛皮による光の減衰を除去するために使用されていました。 in vivoイメージング生体 IM で IVISのスペクトラムを使用して、アルビノC57BL / 6マウスを使用するとシステムの老朽化と、移植の日（キャリパーライフサイエンス、ホプキントン、MA）から可能です。 GL261 - lucおよびGL261 - luc2細胞株は、親のGL261細胞として生体内挙動に同じことを示した。人間性GBMに存在する共有組織学的特徴とこのマウスモデルのようなものがあります：腫瘍壊死、pseudopalisades、血管新生、浸潤、細胞過多、および炎症1。 移植前の動物には、以上のメスで作成された定位装置や切開に入れ、ケタミン（50 mg / kgの）、キシラジン（5 mg / kg）をし、ブプレノルフィン（0.05 mg / kg体重）の腹腔内注射により麻酔し、頭蓋正中線。 burrholeはブレグマ後方0.1ミリメートルと正中線の右側に2.3ミリメートルとした。針は2.6mmの深さに3mmから撤退0.4mmの深さに挿入した。 GL261リュックまたはGL261 - luc2細胞（10 7細胞/ ml）の二つの液を3分間かけて注入した。 burrholeが閉じられましたbonewaxと切開を縫合したと。 定位の移植後の生物の細胞は、移植の日から検出され、腫瘍は、IVISのスペクトラム測定器の3次元画像再構成機能を使用して分析することができます。動物は、150μgルシフェリン/イメージングの前に体重20分の皮下注射を受ける。全身腫瘍組織量が時間の経過とともに平均腫瘍の生物発光を用いて定量される。担癌マウスは、罹患率を評価するために、毎日観察し、以下のいずれかまたは複数の現象が存在するときに安楽死さ：無気力、歩き回るに失敗、猫背の姿勢、新郎への障害、食欲不振、体重の&gt; 10％が失われる。腫瘍は剖検上の動物のすべてに明白であった。

Watch this Scientific Journal Video about Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas at JoVE.com

Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas

C57BL / 6マウスにGL261細胞の頭蓋内注入は、人間の多形性膠芽腫の特徴の多くを再現悪性神経膠腫を生成します。私達は私達が使用できるようにするために安定的にルシフェラーゼを発現するGL261細胞を用い<em&gt; in vivoで</em&gt;イメージングは​​腫瘍の進行に従うこと。手術と3D<em&gt; in vivoで</em&gt;画像が実証されています。

その後の3Dと頭蓋内注入インビボでの</emマウス神経膠腫の>生物発光イメージング

The mouse glioma 261 (GL261) is recognized as an in vivo model system that recapitulates many of the features of human glioblastoma multiforme (GBM). The ...

intracranial-implantation-with-subsequent-3d-vivo-bioluminescent

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas

20.4K ビュー.  Barrow Neurological Institute of St. Joseph’s Hospital and Medical Center. 中文

ビデオ: Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas

Establishing Intracranial Brain Tumor Xenografts With Subsequent Analysis of Tumor Growth and Response to Therapy using Bioluminescence Imaging

Transplantation models using human brain tumor cells have served an essential function in neuro-oncology research for many years. In the past, the most commonly used procedure for human tumor xenograft establishment consisted of the collection of cells from culture flasks, followed by the subcutaneous injection of the collected cells in immunocompromised mice. Whereas this approach still sees frequent use in many laboratories, there has been a significant shift in emphasis over the past decade towards orthotopic xenograft establishment, which, in the instance of brain tumors, requires tumor cell injection into appropriate neuroanatomical structures. Because intracranial xenograft establishment eliminates the ability to monitor tumor growth through direct measurement, such as by use of calipers, the shift in emphasis towards orthotopic brain tumor xenograft models has necessitated the utilization of non-invasive imaging for assessing tumor burden in host animals. Of the currently available imaging methods, bioluminescence monitoring is generally considered to offer the best combination of sensitivity, expediency, and cost. Here, we will demonstrate procedures for orthotopic brain tumor establishment, and for monitoring tumor growth and response to treatment when testing experimental therapies.

Luciferase-modified human brain tumor xenografts can be established intracranially in athymic mice, with subsequent monitoring of tumor growth and response to therapy using bioluminescence imaging. In combination with survival analysis, bioluminescence monitoring is an essential research tool for pre-clinical testing of therapies being considered for treating brain tumors.

生物発光イメージングを使用して腫瘍の成長や治療に対する反応のその後の分析では頭蓋内脳の腫瘍異種移植片の確立

Transplantation models using human brain tumor cells have served an essential function in neuro-oncology research for many years. In the past, the most ...

神経科学

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 ...

医学

Fluorescence Molecular Tomography for In Vivo Imaging of Glioblastoma Xenografts

Tumorigenicity is the capability of cancer cells to form a tumor mass. A widely used approach to determine if the cells are tumorigenic is by injecting immunodeficient mice subcutaneously with cancer cells and measuring the tumor mass after it becomes visible and palpable. Orthotopic injections of cancer cells aim to introduce the xenograft in the microenvironment that most closely resembles the tissue of origin of the tumor being studied. Brain cancer research requires intracranial injection of cancer cells to allow the tumor formation and analysis in the unique microenvironment of the brain. The in vivo imaging of intracranial xenografts monitors instantaneously the tumor mass of orthotopically engrafted mice. Here we report the use of fluorescence molecular tomography (FMT) of brain tumor xenografts. The cancer cells are first transduced with near infrared fluorescent proteins and then injected in the brain of immunocompromised mice. The animals are then scanned to obtain quantitative information about the tumor mass over an extended period of time. Cell pre-labeling allows for cost effective, reproducible, and reliable quantification of the tumor burden within each mouse. We eliminated the need for injecting imaging substrates, and thus reduced the stress on the animals. A limitation of this approach is represented by the inability to detect very small masses; however, it has better resolution for larger masses than other techniques. It can be applied to evaluate the efficacy of a drug treatment or genetic alterations of glioma cell lines and patient-derived samples.

Fluorescence Molecular Tomography for In Vivo Imaging of Glioblastoma Xenografts

Orthotopic intracranial injection of tumor cells has been used in cancer research to study brain tumor biology, progression, evolution, and therapeutic response. Here we present fluorescence molecular tomography of tumor xenografts, which provides real-time intravital imaging and quantification of a tumor mass in preclinical glioblastoma models.

膠芽腫の異種移植片の生体内イメージング用蛍光分子トモグラフィー

Tumorigenicity is the capability of cancer cells to form a tumor mass. A widely used approach to determine if the cells are tumorigenic is by injecting ...

がん研究

Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

Glioblastoma multiforme (GBM) is a high-grade primary brain cancer with a median survival of only 14.6 months in humans despite standard tri-modality treatment consisting of surgical resection, post-operative radiation therapy and temozolomide chemotherapy 1. New therapeutic approaches are clearly needed to improve patient survival and quality of life. The development of more effective treatment strategies would be aided by animal models of GBM that recapitulate human disease yet allow serial imaging to monitor tumor growth and treatment response. In this paper, we describe our technique for the precise stereotactic implantation of bio-imageable GBM cancer cells into the brains of nude mice resulting in tumor xenografts that recapitulate key clinical features of GBM 2. This method yields tumors that are reproducible and are located in precise anatomic locations while allowing in vivo bioluminescent imaging to serially monitor intracranial xenograft growth and response to treatments 3-5. This method is also well-tolerated by the animals with low perioperative morbidity and mortality.

Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

We describe an integrated method for the precise, stereotactic implantation of human glioblastoma multiforme cells into the brains of nude mice and subsequent serial in vivo imaging to monitor growth and response to treatment of the resultant xenografts.

定位頭蓋内注入と<emインビボ&gt;</em多形性膠芽腫のマウスモデル系における腫瘍異種移植片の&gt;生物発光イメージング

Glioblastoma multiforme (GBM) is a high-grade primary brain cancer with a median survival of only 14.6 months in humans despite standard tri-modality ...

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a novel platform that allows for direct long-term visualization of tissue structure changes intracranially. Imaging at a single cell resolution in a real-time fashion provides supplementary dynamic information beyond that provided by standard end-point histological analysis, which looks solely at 'snap-shot' cross sections of tissue.
Establishing this intravital imaging technique in fluorescent chimeric mice, we are able to image four fluorescent channels simultaneously. By incorporating fluorescently labeled cells, such as GFP+ bone marrow, it is possible to track the fate of these cells studying their long-term migration, integration and differentiation within tissue. Further integration of a secondary reporter cell, such as an mCherry glioma tumor line, allows for characterization of cell:cell interactions. Structural changes in the tissue microenvironment can be highlighted through the addition of intra-vital dyes and antibodies, for example CD31 tagged antibodies and Dextran molecules.
Moreover, we describe the combination of our ICW imaging model with a small animal micro-irradiator that provides stereotactic irradiation, creating a platform through which the dynamic tissue changes that occur following the administration of ionizing irradiation can be assessed.
Current limitations of our model include penetrance of the microscope, which is limited to a depth of up to 900 &mu;m from the sub cortical surface, limiting imaging to the dorsal axis of the brain. The presence of the skull bone makes the ICW a more challenging technical procedure, compared to the more established and utilized chamber models currently used to study mammary tissue and fat pads 5-7. In addition, the ICW provides many challenges when optimizing the imaging.

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We describe a novel in vivo imaging technique that couples fluorescent chimeric mice with intracranial windows and high-resolution 2-photon microscopy. This imaging platform aids studies of dynamic changes in brain tissue and microvasculature, at a single-cell level, following pathological insults and is adaptable to assess intracranial drug delivery and distribution.

新規高分解能<em生体内で&gt;</em腫瘍増殖と治療に頭蓋内構造物の動的応答を研究する&gt;イメージング技術

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a ...

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 ...

2P-IVMを用いた脳腫瘍内ナノ粒子のマウスリアルタイム追跡

Two-Photon Intravital Microscopy of Glioblastoma in a Murine Model

The delivery of intravenously administered cancer therapeutics to brain tumors is limited by the blood-brain barrier. A method to directly image the accumulation and distribution of macromolecules in brain tumors in vivo would greatly enhance our ability to understand and optimize drug delivery in preclinical models. This protocol describes a method for real-time in vivo tracking of intravenously administered fluorescent-labeled nanoparticles with two-photon intravital microscopy (2P-IVM) in a mouse model of glioblastoma (GBM).
The protocol contains a multi-step description of the procedure, including anesthesia and analgesia of experimental animals, creating a cranial window, GBM cell implantation, placing a head bar, conducting 2P-IVM studies, and post-surgical care for long-term follow-up studies. We show representative 2P-IVM imaging sessions and image analysis, examine the advantages and disadvantages of this technology, and discuss potential applications.
This method can be easily modified and adapted for different research questions in the field of in vivo preclinical brain imaging.

著者スポットライト:膠芽腫治療における薬物送達と早期発見を最適化するためのマルチモーダルイメージング戦略

We present a novel approach for two-photon microscopy of the tumor delivery of fluorescent-labeled iron oxide nanoparticles to glioblastoma in a mouse model.

マウスモデルにおける神経膠芽腫の2光子生体内顕微鏡

The delivery of intravenously administered cancer therapeutics to brain tumors is limited by the blood-brain barrier. A method to directly image the ...

<meta charset="utf8"></head><body>マウス神経膠芽腫における画像誘導治療送達のための酸化鉄ナノ粒子

Nanoparticle Delivery of an Oligonucleotide Payload in a Glioblastoma Multiforme Animal Model

Glioblastoma multiforme (GBM) is the most common and aggressive form of primary brain malignancy for which there is no cure. The blood-brain barrier is a significant hurdle in the delivery of therapies to GBM. Reported here is an image-guided, iron oxide-based therapeutic delivery nano platform capable of bypassing this physiological barrier by virtue of size and accumulating in the tumor region, delivering its payload. This 25 nm nano platform consists of crosslinked dextran-coated iron oxide nanoparticles labeled with Cy5.5 fluorescent dye and containing antisense oligonucleotide as a payload. The magnetic iron oxide core enables tracking of the nanoparticles through in vivo magnetic resonance imaging, while Cy5.5 dye allows tracking by optical imaging. This report details the monitoring of the accumulation of this nanoparticle platform (termed MN-anti-miR10b) in orthotopically implanted glioblastoma tumors following intravenous injection. In addition, it provides insight into the in vivo delivery of RNA oligonucleotides, a problem that has hampered the translation of RNA therapeutics into the clinic.

<meta charset="utf8"></head><body>著者スポットライト:膠芽腫治療のための酸化鉄ナノ粒子による革新的ながん治療

The blood-brain barrier is a significant hurdle in the delivery of therapies for glioblastoma, a disease for which there is no cure. Here, we report an in vivo image-guided iron oxide therapeutic nano platform that can bypass this physiological barrier by virtue of size and accumulate in the tumor.

神経膠芽腫多形動物モデルにおけるオリゴヌクレオチドペイロードのナノ粒子送達

Glioblastoma multiforme (GBM) is the most common and aggressive form of primary brain malignancy for which there is no cure. The blood-brain barrier is a ...

その後の3Dと頭蓋内注入インビボでの生物発光イメージング

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生物発光イメージングを使用して腫瘍の成長や治療に対する反応のその後の分析では頭蓋内脳の腫瘍異種移植片の確立

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

膠芽腫の異種移植片の生体内イメージング用蛍光分子トモグラフィー

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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

マウスモデルにおける神経膠芽腫の2光子生体内顕微鏡

マウスモデルにおける神経膠芽腫の2光子生体内顕微鏡

マウスモデルにおける神経膠芽腫の2光子生体内顕微鏡

神経膠芽腫多形動物モデルにおけるオリゴヌクレオチドペイロードのナノ粒子送達