Name: chemotherapy-induced vascular toxicity - real-time in vivo imaging of vessel impairment
Uploaded: 2015-01-07T13:00:00.000+00:00
Duration: 4 min 48 s
Description: Watch this Scientific Journal Video about Chemotherapy-induced Vascular Toxicity - Real-time In vivo Imaging of Vessel Impairment at JoVE.com

Ethics Statement: Alle Experimente wurden von der Institutional Animal Care und Verwenden Committee genehmigt. Tierpflege war gemäß gültiger Richtlinien. ICR weibliche Mäuse (7-8 Wochen alt, 25 bis 30 g) wurden in Klimaanlage, Licht gesteuert Tiereinrichtungen des Sackler Fakultät für Medizin in Tel-Aviv Universität untergebracht. Am Begriff wurden die Tiere mit Narkosedosis euthanasiert. 1. Fibred konfokale Fluoreszenzmikroskopie (FCFM) Kalibrierung <ol><li> Schalten Sie das Gerät EIN. </li><li> Schließen Sie die Mikrosonde (mini0 / 30). </li><li> Kalibrieren der Vorrichtung nach den Anweisungen des Herstellers. </li></ol> 2. Mäuse Vorbereitung für Imaging <ol><li> Anesthetize durch eine subkutane Injektion von sowohl Ketaset (100 mg / kg) und XYL-M2 (6 mg / kg). Überprüfen Sie die Betäubung durch Teilnahmslosigkeit bis Fuß-Prise. </li><li> Inzision der Haut unter der Leiste, um<html> zeigen die Oberschenkel arteriellen Gefäße. Halten Sie die Inzisionsstelle feucht mit Kochsalzlösung nach Einschnitt. </li><li> Heizen Sie den Schwanz durch die Verwendung eines Beutels (oder einen Handschuh) mit warmem Wasser (nicht zu heiß zum Anfassen) für ca. 30 sec gefüllt. Vorbereiten eines intravenösen (IV) Shunt zur Verabreichung von FITC-Dextran (ein Kontrastmittel) und entweder Kochsalzlösung oder chemotherapeutisches Mittel, durch Einführen einer Nadel (30 G, 1/2 Zoll) in die Schwanzvene und Anbringen einer 1 ml Spritze, um es . Achten Sie darauf, die Ader ist durch die Injektion von Kochsalzlösung auf. Hinweis: eine IV-Verabreichung von FITC-Dextran (mit hohem Molekulargewicht; 100 &amp; mgr; l; 10 mg / ml; 2.000 kDa) erleichtert die Visualisierung des Oberschenkel Mikrovaskulatur von FCFM. Doxorubicin (100 ul, 8 mg / kg Adriamycin) oder Kochsalzlösung wird ebenfalls später verabreicht werden IV in die vorgewärmte Schwanzvene. </li><li> Bewegen Sie die Maus supinely auf einer Polystyrol-Bühne. Sichern Sie die Maus auf dem Pad und zu pflegen, die mithilfe von chirurgischen Klebeband. </li></ol>title &quot;&gt; 3. Abbildung von Oberschenkelblutgefäße durch FCFM während und nach der Verabreichung von Doxorubicin oder Saline Anmerkung: (1) Mikrosonde (mini0 / 30): Die Fasern versehenen Konfokalmikroskop in dieser Studie verwendet wird, aus zwei Einheiten zusammengesetzt ist. (2) Laserscaneinheit (LSU-488; 488 nm Wellenlänge). <ol><li> Führen Sie alle Zeit-Runden-Analysen unter Verwendung der LSU 488 nm Wellenlängen-Laser. HINWEIS: Das Hauptgerät Detektor die gefiltert (500 bis 650 nm) emittierte Fluoreszenz. Die aufgenommenen Bilder werden anschließend wieder aufgebaut und mit einer Geschwindigkeit von 12 Bildern / s angezeigt. </li><li> Trennen Sie vorsichtig die Spritze von der Nadel und fügen Sie eine neue Spritze mit FITC-Dextran. Verwalten (IV) 100 ul FITC-Dextran. </li><li> Verschieben Sie die Mikrosonde (mini0 / 30) an eine geeignete Sichtfeld und fixiert es, nach Anpassung an die z-Achse, um das entsprechende Bild zu erhalten. Warten Sie auf das Anfangssignal zu verblassen, bis eine klare und fokussierte Signal visible. </li><li> Nehmen Sie eine Grundlinie Blutfluss für kurze Stabilisierungsphase (~ 30 sec). Schließen Sie dann auf die Nadel eine andere Spritze, die entweder Doxorubicin oder Kochsalzlösung. IV verwalten 100 ul Doxorubicin oder Kochsalzlösung. </li><li> Überwachung der Strömung des eingespritzten FITC-Dextran kontinuierlich für 20 min. Auf der FCFM-zugehörige Software, verwenden Sie die Schaltfläche Durchmesser am oberen Lineal, um die Blutgefäße zu messen und kategorisieren sie als kleine (&lt;15 um) oder große (&gt; 15 um). </li><li> Euthanize das Tier mit Narkosedosis. </li></ol>

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	<li>Chow, A. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anthracyclines+cause+endothelial+injury+in+pediatric+cancer+patients:+a+pilot+study.">Anthracyclines cause endothelial injury in pediatric cancer patients: a pilot study.</a> J Clin Oncol. 24 (6), 925-928 (2006).</li><li>Nuver, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+chemotherapy-induced+cardiovascular+changes+in+patients+with+testicular+cancer.">Acute chemotherapy-induced cardiovascular changes in patients with testicular cancer.</a> J Clin Oncol. 23 (36), 9130-9137 (2005).</li><li>Vos, F. 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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+tissue+factor+on+human+monocytes+by+cisplatin+and+adriamycin.">Modulation of tissue factor on human monocytes by cisplatin and adriamycin.</a> Br J Haematol. 81 (4), 480-488 (1992).</li><li>Kotamraju, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Doxorubicin-induced+apoptosis+in+endothelial+cells+and+cardiomyocytes+is+ameliorated+by+nitrone+spin+traps+and+ebselen.+Role+of+reactive+oxygen+and+nitrogen+species.">Doxorubicin-induced apoptosis in endothelial cells and cardiomyocytes is ameliorated by nitrone spin traps and ebselen. 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Auswerten Chemotherapie-induzierter vaskulärer Toxizität stellt aufgrund der Schwierigkeit bei der Visualisierung der Dynamik der Vaskulatur in Antwort auf Stimuli in Echtzeit. Zahlreiche klinische Studien haben impliziert, dass mehrere Chemotherapien verursachen direkte Gefäßverletzung, aber der Mechanismus und Besonderheiten dieser Toxizität ist noch nicht geklärt. Wir haben eine Echtzeit für die Bewertung der potentiellen Gefäß Toxizität der Chemotherapie in Mäusen, umfassend von gefaserten konfokale Fluor...

Klinische Daten zeigen, dass verschiedene Klassen von Chemotherapien zu entlocken eine Vielzahl von Gefäßerkrankungen von Raynaud-Phänomen, Bluthochdruck, myocardialinfarction, zerebrovaskuläre Angriff und Lebervenen occlusivedisease 1,2 manifestiert. &quot;&#39;Unbeabsichtigter&#39; anti-angiogenen Drogen&quot; ist ein recht neuer Begriff, der herkömmliche Chemotherapeutika, die als mögliche Angiogenese-Inhibitoren wirken beschreibt, obwohl sie ursprünglich nicht für diesen Zweck entwickelt, aber 3-5 entwickelt, um Tumorzellen zu eliminieren durch Auferlegung so wenig &quot;Sicherheiten Schaden &quot;zu normalen Zellen wie möglich 3. Mehrere Chemotherapien wurden als vasculo-Giftstoffe angedeutet worden, wie in klinischen Studien mit Serum-Biomarkern beobachtet. Unter diesen sind Alkylierungsmittel (wie Cyclophosphamid), Platin-Verbindungen (wie Cisplatin) und Anthrazyklinen 1,2,5-7. Akute kardiovaskulären Komplikationen können als resul auftretent von vaskulären Toxizität durch Chemotherapie. Sie können in chronischen Erkrankungen wie Arteriosklerose und machen erhöhtes Risiko für Spät vaskulären Morbidität Fortschritt. Trotz Montage klinische Beweise, gibt es einen Mangel an bezeichneten Studien, in der der Mechanismus der Gefäß Toxizität und daher ist eine weitere Erläuterung der genaue Pathogenese sie anrichten gerechtfertigt. Eine große Herausforderung bei der Aufdeckung des Mechanismus der Chemotherapie-induzierter vaskulärer Toxizität ergibt sich aus der Komplexität der Untersuchung von Gefäßfunktion in vivo. Wir beschreiben hier eine Plattform für hochauflösende in vivo molekulare Bildgebung bei Mäusen, die den Blutfluss und Schiffe Eigenschaften erfassen können. Diese Plattform erleichtert den Nachweis der direkten Behandlung induzierten vaskulären Wirkungen: in Echtzeit, sowie die folgenden sie über eine Zeitdauer, innerhalb der gleichen Individuen.

<table><tbody><tr><td>general anesthesia</td><td>Fort Dodge Animal Health, IA, USA and Biove Laboratories, France</td><td></td><td>100 mg/kg ketaset and 6 mg/kg XYL-M2</td></tr><tr><td>depilatory cream (Veet)</td><td>ReckittBenckiser, Bristol, UK</td><td></td><td></td></tr><tr><td>30 G, 1/2 inch needle attached to 1 ml syringe</td><td></td><td></td><td></td></tr><tr><td>FITC dextran (10 mg/ml; MW 2,000 kDa)</td><td>Sigma</td><td>FD2000S</td><td>100 &amp;mu;l volume</td></tr><tr><td>Doxorubicin</td><td>Teva, Israel</td><td></td><td>8 mg/kg, Adriamycin</td></tr><tr><td>paclitaxel</td><td>Taro, Israel</td><td></td><td>1.2 mg/kg, Medexel</td></tr><tr><td>saline</td><td></td><td></td><td></td></tr></tbody></table>

chemotherapy-induced vascular toxicity - real-time in vivo imaging of vessel impairment

Bestimmte Arten von Chemotherapien können akute Gefäßveränderungen, die in langfristigen Bedingungen, die den Patienten zu einem erhöhten Risiko von vaskulären Erkrankungen begünstigen können, fortschreiten kann auszuüben. Doch wenn auch der Befestigungs klinische Beweise, gibt es einen Mangel an klaren Studien des vaskulären Toxizität und daher der Ätiologie einer heterogenen Gruppe von Gefäß / kardiovaskulären Störungen noch aufgeklärt werden. Darüber hinaus kann der Mechanismus, der vaskulären Toxizität zugrunde liegen vollständig unterscheiden sich von den Grundsätzen der Chemotherapie-induzierter Kardiotoxizität, die in Beziehung steht zu Myozyten Verletzungen zu lenken. Wir haben eine Echtzeit eingerichtet, in vivo molekularen Bildgebung Plattform, um die mögliche akute vaskuläre Toxizität von Antikrebstherapien zu bewerten. Wir haben eine Plattform für in vivo, hochauflösende molekulare Bildgebung bei Mäusen, die für die Visualisierung Gefäßsystem im geschlossenen Organe und Referenz blo eingestelltod Gefäße innerhalb derselben Personen während jedes einzelnen dienen als seine eigene Kontrolle. Wände der Blutgefäße wurden nach Doxorubicin Verwaltung beeinträchtigt, was einen einzigartigen Mechanismus von vaskulären Toxizität, die der frühes Ereignis in der Endorgane Verletzung sein kann. Hierbei wird das Verfahren zum fibered konfokale Fluoreszenzmikroskopie (FCFM) basierend Abbildungs ​​beschrieben, die einen innovativen Modus bietet den physiologischen Phänomenen auf zellulärer und subzellulärer in Tier Themen zu verstehen.

In-vivo-Bildgebung bei kontinuierlichen Echtzeit- Die bildgebende Vorrichtung verwendet, hier ist ein hochauflösendes, fibered konfokalen Mikroskop, mit einer Sonde, die Visualisierung des Gefäßsystems und ihre Reaktion auf verschiedene Stimuli, wie Chemotherapie ermöglicht. Dieses Verfahren ist minimal-invasiv, da, obwohl es Bildgebung der tiefen Gefäße oder Organe zu erleichtern, einen kleinen Einschnitt für die Sonde erfordert. Die Sondenbündel bestehen aus Zehntausenden von...

Watch this Scientific Journal Video about Chemotherapy-induced Vascular Toxicity - Real-time In vivo Imaging of Vessel Impairment at JoVE.com

Chemotherapy-induced Vascular Toxicity - Real-time In vivo Imaging of Vessel Impairment

We herein describe the method of fibered confocal fluorescent microscopy (FCFM) based imaging, which provides an innovative mode to understand physiological phenomena at the cellular and sub-cellular levels in animal subjects.

Chemotherapie-induzierte vaskuläre Toxizität - Echtzeit-<em&gt; In-vivo-</em&gt; Imaging von Vessel Impairment

Certain classes of chemotherapies may exert acute vascular changes that may progress into long-term conditions that may predispose the patient to an ...

chemotherapy-induced-vascular-toxicity-real-time-vivo-imaging-vessel

Research

JoVE Journal

Medicine

Chemotherapy-induced Vascular Toxicity - Real-time In vivo Imaging of Vessel Impairment

7.3K Aufrufe.  Tel Aviv University. We herein describe the method of fibered confocal fluorescent microscopy (FCFM) based imaging, which provides an innovative mode to understand physiological phenomena at the cellular and sub-cellular levels in animal subjects.

Serie: Chemotherapy-induced Vascular Toxicity - Real-time In vivo Imaging of Vessel Impairment

Contrast Enhanced Vessel Imaging using MicroCT

Microscopic computed tomography (microCT) offers high-resolution volumetric imaging of the anatomy of living small animals. However, the contrast between different soft tissues and body fluids is inherently poor in micro-CT images 1. Under these circumstances, visualization of blood vessels becomes a nearly impossible task. To overcome this and to improve the visualization of blood vessels exogenous contrast agents can be used. Herein, we present a methodology for visualizing the vascular network in a rodent model. By using a long-acting aqueous colloidal polydisperse iodinated blood-pool contrast agent, eXIA 160XL, we optimized image acquisition parameters and volume-rendering techniques for finding blood vessels in live animals. Our findings suggest that, to achieve a superior contrast between bone and soft tissue from vessel, multiple-frames (at least 5-8/ frames per view), and 360-720 views (for a full 360&deg; rotation) acquisitions were mandatory. We have also demonstrated the use of a two-dimensional transfer function (where voxel color and opacity was assigned in proportion to CT value and gradient magnitude), in visualizing the anatomy and highlighting the structure of interest, the blood vessel network. This promising work lays a foundation for the qualitative and quantitative assessment of anti-angiogenesis preclinical studies using transgenic or xenograft tumor-bearing mice.

Contrast enhanced small animal vessel imaging by microCT is a rapid, cost-effective and high-throughput technique for serial in situ examination for tumor development, for analyzing the network of blood vessels that nourish them, and for following the response of tumors to preclinical therapeutic intervention(s).

Kontrastverstärkten Vessel Imaging mit MicroCT

Microscopic computed tomography (microCT) offers high-resolution volumetric imaging of the anatomy of living small animals. However, the contrast between ...

Forschung

Medizin

Live Imaging of Drug Responses in the Tumor Microenvironment in Mouse Models of Breast Cancer

The tumor microenvironment plays a pivotal role in tumor initiation, progression, metastasis, and the response to anti-cancer therapies. Three-dimensional co-culture systems are frequently used to explicate tumor-stroma interactions, including their role in drug responses. However, many of the interactions that occur in vivo in the intact microenvironment cannot be completely replicated in these in vitro settings. Thus, direct visualization of these processes in real-time has become an important tool in understanding tumor responses to therapies and identifying the interactions between cancer cells and the stroma that can influence these responses. Here we provide a method for using spinning disk confocal microscopy of live, anesthetized mice to directly observe drug distribution, cancer cell responses and changes in tumor-stroma interactions following administration of systemic therapy in breast cancer models. We describe procedures for labeling different tumor components, treatment of animals for observing therapeutic responses, and the surgical procedure for exposing tumor tissues for imaging up to 40 hours. The results obtained from this protocol are time-lapse movies, in which such processes as drug infiltration, cancer cell death and stromal cell migration can be evaluated using image analysis software.

We describe a method for imaging response to anti-cancer treatment in vivo and at single cell resolution.

Live-Imaging of Drug Responses in der Tumor-Mikroumgebung in Mausmodellen von Brustkrebs

The tumor microenvironment plays a pivotal role in tumor initiation, progression, metastasis, and the response to anti-cancer therapies. Three-dimensional ...

Real-time Imaging of Heterotypic Platelet-neutrophil Interactions on the Activated Endothelium During Vascular Inflammation and Thrombus Formation in Live Mice

Interaction of activated platelets and leukocytes (mainly neutrophils) on the activated endothelium mediates thrombosis and vascular inflammation.1,2 During thrombus formation at the site of arteriolar injury, platelets adherent to the activated endothelium and subendothelial matrix proteins support neutrophil rolling and adhesion.3 Conversely, under venular inflammatory conditions, neutrophils adherent to the activated endothelium can support adhesion and accumulation of circulating platelets. Heterotypic platelet-neutrophil aggregation requires sequential processes by the specific receptor-counter receptor interactions between cells.4 It is known that activated endothelial cells release adhesion molecules such as von Willebrand factor, thereby initiating platelet adhesion and accumulation under high shear conditions.5 Also, activated endothelial cells support neutrophil rolling and adhesion by expressing selectins and intercellular adhesion molecule-1 (ICAM-1), respectively, under low shear conditions.4 Platelet P-selectin interacts with neutrophils through P-selectin glycoprotein ligand-1 (PSGL-1), thereby inducing activation of neutrophil &beta;2 integrins and firm adhesion between two cell types. Despite the advances in in vitro experiments in which heterotypic platelet-neutrophil interactions are determined in whole blood or isolated cells,6,7 those studies cannot manipulate oxidant stress conditions during vascular disease. In this report, using fluorescently-labeled, specific antibodies against a mouse platelet and neutrophil marker, we describe a detailed intravital microscopic protocol to monitor heterotypic interactions of platelets and neutrophils on the activated endothelium during TNF-&alpha;-induced inflammation or following laser-induced injury in cremaster muscle microvessels of live mice.

Here we report an experimental technique of fluorescence intravital microscopy to visualize heterotypic platelet-neutrophil interactions on the activated endothelium during vascular inflammation and thrombus formation in live mice. This microscopic technology will be valuable to study the molecular mechanism of vascular disease and to test pharmacologic agents under pathophysiological conditions.

Real-time Imaging der heterotypische Platelet-Neutrophilen-Interaktionen auf aktiviertem Endothel Während Vascular Entzündung und Thrombusbildung in lebenden Mäusen

Interaction of activated platelets and leukocytes (mainly neutrophils) on the activated endothelium mediates thrombosis and vascular inflammation.1,2 ...

Immunologie und Infektion

In vivo Imaging of Tumor Angiogenesis using Fluorescence Confocal Videomicroscopy

Fibered confocal fluorescence in vivo imaging with a fiber optic bundle uses the same principle as fluorescent confocal microscopy. It can excite fluorescent in situ elements through the optical fibers, and then record some of the emitted photons, via the same optical fibers. The light source is a laser that sends the exciting light through an element within the fiber bundle and as it scans over the sample, recreates an image pixel by pixel. As this scan is very fast, by combining it with dedicated image processing software, images in real time with a frequency of 12 frames/sec can be obtained.
We developed a technique to quantitatively characterize capillary morphology and function, using a confocal fluorescence videomicroscopy device. The first step in our experiment was to record 5 sec movies in the four quadrants of the tumor to visualize the capillary network. All movies were processed using software (ImageCell, Mauna Kea Technology, Paris France) that performs an automated segmentation of vessels around a chosen diameter (10 &mu;m in our case). Thus, we could quantify the 'functional capillary density', which is the ratio between the total vessel area and the total area of the image. This parameter was a surrogate marker for microvascular density, usually measured using pathology tools.
The second step was to record movies of the tumor over 20 min to quantify leakage of the macromolecular contrast agent through the capillary wall into the interstitium. By measuring the ratio of signal intensity in the interstitium over that in the vessels, an 'index leakage' was obtained, acting as a surrogate marker for capillary permeability.

In vivo Imaging of Tumor Angiogenesis using Fluorescence Confocal Videomicroscopy

In this paper, we present a method to analyze tumor microvessels in vivo using dynamic contrast-enhanced fluorescence videomicroscopy. Two quantitative parameters were acquired: functional capillary density reflecting the vascularity of the tumor, and index leakage reflecting the leakiness of the endothelial wall.

In-vivo-Imaging der Tumorangiogenese mit Fluoreszenz-Videomikroskopie Konfokale

Fibered confocal fluorescence in vivo imaging with a fiber optic bundle uses the same principle as fluorescent confocal microscopy. It can excite ...

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.

A Novel Hochauflösende<em&gt; In vivo</em&gt; Bildtechnik, um die Dynamik der intrakranielle Strukturen zu Tumorwachstum und Therapeutics Studieren

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

Erstellung eines mikrofluidikbasierten vaskularisierten humanen Mikrotumormodells

Establishing a Physiologic Human Vascularized Micro-Tumor Model for Cancer Research

A lack of validated cancer models that recapitulate the tumor microenvironment of solid cancers in vitro remains a significant bottleneck for preclinical cancer research and therapeutic development.&#160;To overcome this problem, we have developed the vascularized microtumor (VMT), or tumor chip, a microphysiological system that realistically models the complex human tumor microenvironment. The VMT forms de novo within a microfluidic platform by co-culture of multiple human cell types under dynamic, physiological flow conditions. This tissue-engineered micro-tumor construct incorporates a living perfused vascular network that supports the growing tumor mass just as newly formed vessels do in vivo. Importantly, drugs and immune cells must cross the endothelial layer to reach the tumor, modeling in vivo physiological barriers to therapeutic delivery and efficacy. Since the VMT platform is optically transparent, high-resolution imaging of dynamic processes such as immune cell extravasation and metastasis can be achieved with direct visualization of fluorescently labeled cells within the tissue. Further, the VMT retains in vivo tumor heterogeneity, gene expression signatures, and drug responses. Virtually any tumor type can be adapted to the platform, and primary cells from fresh surgical tissues grow and respond to drug treatment in the VMT, paving the way toward truly personalized medicine. Here, the methods for establishing the VMT and utilizing it for oncology research are outlined. This innovative approach opens new possibilities for studying tumors and drug responses, providing researchers with a powerful tool to advance cancer research.

Autor im Rampenlicht: Schaffung humaner vaskularisierter Mikrotumoren als Modelle für die translationale Krebsforschung 

This protocol presents a physiologically relevant tumor-on-a-chip model to perform high-throughput basic and translational human cancer research, advancing drug screening, disease modeling, and personalized medicine approaches with a description of loading, maintenance, and evaluation procedures.

Etablierung eines physiologischen humanen vaskularisierten Mikrotumormodells für die Krebsforschung

A lack of validated cancer models that recapitulate the tumor microenvironment of solid cancers in vitro remains a significant bottleneck for preclinical ...

Krebsforschung

Geführte Tumorinokulation und multimodale Bildgebung bei Mäusen

A Dorsal Skinfold Window Chamber Tumor Mouse Model for Combined Intravital Microscopy and Magnetic Resonance Imaging in Translational Cancer Research

Preclinical intravital imaging such as microscopy and optical coherence tomography have proven to be valuable tools in cancer research for visualizing the tumor microenvironment and its response to therapy. These imaging modalities have micron-scale resolution but have limited use in the clinic due to their shallow penetration depth into tissue. More clinically applicable imaging modalities such as CT, MRI, and PET have much greater penetration depth but have comparatively lower spatial resolution (mm scale). 
To translate preclinical intravital imaging findings into the clinic, new methods must be developed to bridge this micro-to-macro resolution gap. Here we describe a dorsal skinfold window chamber tumor mouse model designed to enable preclinical intravital and clinically applicable (CT and MR) imaging in the same animal, and the image analysis platform that links these two disparate visualization methods. Importantly, the described window chamber approach enables the different imaging modalities to be co-registered in 3D using fiducial markers on the window chamber for direct spatial concordance. This model can be used for validation of existing clinical imaging methods, as well as for the development of new ones through direct correlation with "ground truth" high-resolution intravital findings. 
Finally, the tumor response to various treatments-chemotherapy, radiotherapy, photodynamic therapy-can be monitored longitudinally with this methodology using preclinical and clinically applicable imaging modalities. The dorsal skinfold window chamber tumor mouse model and imaging platforms described here can thus be used in a variety of cancer research studies, for example, in translating preclinical intravital microscopy findings to more clinically applicable imaging modalities such as CT or MRI.

Autor im Rampenlicht: Integration von hochauflösender Intravitalbildgebung und MRT zur Verbesserung der Planung der stereotaktischen Körperstrahlentherapie

Translation of Intravital microscopy findings is challenged by its shallow depth penetration into tissue. Here we describe a dorsal window chamber mouse model that enables co-registration of intravital microscopy and clinically applicable imaging modalities (e.g., CT, MRI) for direct spatial correlation, potentially streamlining clinical translation of intravital microscopy findings.

Ein Mausmodell eines dorsalen Hautfaltenfensterkammertumors für die kombinierte Intravitalmikroskopie und Magnetresonanztomographie in der translationalen Krebsforschung

Preclinical intravital imaging such as microscopy and optical coherence tomography have proven to be valuable tools in cancer research for visualizing the ...

Chemotherapie-induzierte vaskuläre Toxizität - Echtzeit-

Zusammenfassung

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Etablierung eines physiologischen humanen vaskularisierten Mikrotumormodells für die Krebsforschung

Etablierung eines physiologischen humanen vaskularisierten Mikrotumormodells für die Krebsforschung

Etablierung eines physiologischen humanen vaskularisierten Mikrotumormodells für die Krebsforschung

Ein Mausmodell eines dorsalen Hautfaltenfensterkammertumors für die kombinierte Intravitalmikroskopie und Magnetresonanztomographie in der translationalen Krebsforschung

Ein Mausmodell eines dorsalen Hautfaltenfensterkammertumors für die kombinierte Intravitalmikroskopie und Magnetresonanztomographie in der translationalen Krebsforschung