伦理学声明:所有实验均批准的机构动物护理和使用委员会。根据机构指引照顾动物了。 ICR雌性小鼠（7 - 8周龄，25 - 30克）被安置在了空调，光控动物特拉维夫大学医学系赛克勒设施。在术语，动物安乐死麻醉过量。 1. Fibred共聚焦荧光显微镜（FCFM）校准<ol><li>打开该设备。 </li><li>连接探针（mini0 / 30）。 </li><li>根据制造商的说明校准装置。 </li></ol> 2.准备小鼠成像<ol><li>通过皮下注射既Ketaset（100毫克/千克）和XYL-M2（6毫克/千克）的麻醉。确定合适的麻醉通过反应迟钝到脚趾捏。 </li><li>切开腹股沟皮肤下面以<html>揭示了股动脉血管。请跟随切口切口部位湿润用生理盐水。 </li><li>用袋（或手套）约30秒盛有温水（不烫手）加热尾巴。制备的静脉内（IV）的分流为FITC葡聚糖（造影剂）的施用与盐水或化疗剂，通过插入一针（30克，1/2英寸）至尾静脉，并附加一个1ml注射器将其。确保静脉注射生理盐水打开。 注:FITC葡聚糖的IV施用（高分子量; 100微升; 10毫克/毫升; 2000 kDa的）便于可视化股骨微血管由FCFM。阿霉素（100微升; 8毫克/公斤，阿霉素），或盐水也将在后面IV施用到预加热的尾静脉。 </li><li>仰起将鼠标上的聚苯乙烯阶段。确保鼠标垫，用手术胶带保持位置。 </li></ol>标题"&gt; 3。显像股血管通过FCFM在和阿霉素或生理盐水给药后注意:在此研究中使用的纤维质共聚焦显微镜包括两个单元:（1）微探针（mini0 / 30）。 （2）激光扫描单元（LSU-488; 488nm的波长下）。 <ol><li>执行所有时间圈分析使用LSU 488纳米波长的激光。 注:主单元探测器探测过滤（500 - 650纳米）发出荧光。所获取的图像被事后重建，并在12帧/秒的速率显示。 </li><li>小心地断开从针的注射器，并附加包含FITC标记的葡聚糖的新注射器。管理（IV）100微升FITC右旋糖酐。 </li><li>移的探针（mini0 / 30），视合适字段和固定它，以下调整z轴，以获得相应的图像。等待开始信号淡出直到一个明确的和聚焦的信号是VISIBLE。 </li><li>记录基线血流短暂的稳定期（〜30秒）。然后连接到另一针注射器，要么含有阿霉素或生理盐水。四，管理100微升阿霉素或生理盐水。 </li><li>监视注入FITC-葡聚糖的流量连续地进行20分钟。在FCFM相关软件，使用直径按钮的上标尺以测量血管和它们分类为小（&lt;15微米）或大（&gt; 15微米）。 </li><li>安乐死麻醉过量的动物。 </li></ol>

<ol>
	<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. 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=Endothelial+cell+effects+of+cytotoxics:+balance+between+desired+and+unwanted+effects.">Endothelial cell effects of cytotoxics: balance between desired and unwanted effects.</a> Cancer Treat Rev. 30 (6), 495-513 (2004).</li><li>Kerbel, R. 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='Accidental'+anti-angiogenic+drugs.+anti-oncogene+directed+signal+transduction+inhibitors+and+conventional+chemotherapeutic+agents+as+examples.Eur.">'Accidental' anti-angiogenic drugs. anti-oncogene directed signal transduction inhibitors and conventional chemotherapeutic agents as examples.Eur.</a> J Cancer. 36 (10), 1248-1257 (2000).</li><li>Soultati, A., 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=Endothelial+vascular+toxicity+from+chemotherapeutic+agents:+preclinical+evidence+and+clinical+implications.">Endothelial vascular toxicity from chemotherapeutic agents: preclinical evidence and clinical implications.</a> Cancer Treat Rev. 38 (5), 473-483 (2012).</li><li>Tempelhoff, G. F., 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=Blood+coagulation+during+adjuvant+epirubicin/cyclophosphamide+chemotherapy+in+patients+with+primary+operable+breast+cancer.">Blood coagulation during adjuvant epirubicin/cyclophosphamide chemotherapy in patients with primary operable breast cancer.</a> J Clin Oncol. 14 (9), 2560-2568 (1996).</li><li>Ben Aharon, I., 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+vascular+toxicity--targeting+potential+pathways+may+reduce+procoagulant+activity.">Doxorubicin-induced vascular toxicity--targeting potential pathways may reduce procoagulant activity.</a> PLoS One. 8 (9), e7515 (2013).</li><li>Laemmel, E., 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=Fibered+confocal+fluorescence+microscopy+(Cell-viZio)+facilitates+extended+imaging+in+the+field+of+microcirculation.+A+comparison+with+intravital+microscopy.">Fibered confocal fluorescence microscopy (Cell-viZio) facilitates extended imaging in the field of microcirculation. A comparison with intravital microscopy.</a> J Vasc Res. 41 (5), 400-411 (2004).</li><li>Al-Gubory, K. H., Houdebine, L. 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P., Mehta, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+toxicity+of+anthracyclines+on+cultured+endothelial+cells.">Differential toxicity of anthracyclines on cultured endothelial cells.</a> Endothelium. 11 (5-6), 253-258 (2004).</li><li>Kim, E. 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=Doxorubicin-induced+platelet+cytotoxicity:+a+new+contributory+factor+for+doxorubicin-mediated+thrombocytopenia.">Doxorubicin-induced platelet cytotoxicity: a new contributory factor for doxorubicin-mediated thrombocytopenia.</a> J Thromb Haemost. 7 (7), 1172-1183 (2009).</li><li>Walsh, J., Wheeler, H. R., Geczy, C. 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. Role of reactive oxygen and nitrogen species.</a> J Biol Chem. 275 (43), 33585-33592 (2000).</li><li>Vasquez-Vivar, 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=Endothelial+nitric+oxide+synthase-dependent+superoxide+generation+from+adriamycin.">Endothelial nitric oxide synthase-dependent superoxide generation from adriamycin.</a> Biochemistry. 36 (38), 11293-11297 (1997).</li></ol>

评价化疗诱导的血管毒性是具有挑战性，因为在可视化血管的动态响应于实时一个刺激的难度。大量临床研究已经牵连的几个化疗造成直接损伤血管，但这种毒性的机制和特点仍有待澄清。我们建立了实时， 体内分子成像的平台如本文8-10描述评价化疗在含有纤维状共聚焦荧光显微镜的小鼠的潜在血管毒性。这种高解析度的分子的小鼠成像是适合用于可视化动脉血流和血管&#39;架构?...

临床证据表明，几类化疗引起的各种由雷诺现象，高血压，心肌梗死，脑血管攻击，肝静脉occlusivedisease 1,2体现血管病症。 "&#39;意外&#39;抗血管生成药物"是一个相当新的名词，它描述了传统的化疗药物是作为可能的血管生成抑制剂，虽然他们不是最初开发用于这一目的3-5，但设计的征收尽可能少的消除肿瘤细胞的"抵押品损害"正常细胞尽可能3。几种化疗已经暗示为血管 - 毒物在应用血清生物标记物的临床研究中观察到。在这些被烷化剂（如环磷酰胺），铂化合物（例如顺铂）和蒽环类1,2,5-7。 急性心脑血管并发症，可能会出现一个resul吨血管毒性化疗所致的。他们可能会进步到像动脉粥样硬化，占后期血管发病的危险性增加慢性疾病。然而，尽管安装的临床证据，有指定的研究强调血管毒性的机制的缺乏，因此，它们造成的确切发病机理的进一步澄清是必要的。 在揭示化疗引起的血管毒性机制的一个主要挑战来源于体内调查血管功能的复杂性。我们描述了本文中的高清晰度的体内分子成像的小鼠，使捕捉血流和血管&#39;特性的一个平台。这个平台便于直接治疗性血管效应的检测:实时，以及跟随它们过了一段时间的同一个体中。

<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

化学疗法的某些类可能施加急性血管变化，可能进展成长期条件可能会使病人血管并发症的风险增加。然而，尽管所述安装临床证据，有血管毒性，因此，血管/心血管病症一组异质性的病因的明确研究的缺乏尚待阐明。此外，该可依据血管毒性的机制可以完全由化疗引起的心脏毒性，这是直接细胞损伤相关的原理不同。我们建立了实时， 体内分子成像平台以评价抗癌疗法的潜在急性血管毒性。 我们已经建立了体内 ，小鼠的高分辨率分子成像，一个平台适用于密闭机关和参照一个BLO内血管可视化而每个单独的同一个体内外径船只充当其自身的控制。阿霉素给药后的血管壁均受损，代表血管毒性的一个独特的机制，可能是在终末器官损伤的早期事件。在此，纤维状共聚焦荧光显微镜（FCFM）基于成像的方法进行说明，它提供了一个创新的方式来了解生理现象在细胞和亚细胞水平的动物受试者。

体内连续成像在实时此处所使用的成像装置是一个高清晰度，纤维状共焦显微镜，装备有探针，使脉管的可视化和其响应于各种刺激如化疗。这种方法是微创因为尽管它可以便于深血管或器官的成像，它需要一个小切口的探针。探头束组成的纤维，光学显微镜和专有精密连接器数万。的示意图示于图1。 股骨头微血管成像<p class="jove_conte...

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.

化疗引起的血管毒性 - 实时<em&gt;在体内</em&gt;船舶减值成像

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

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

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

7.4K 次观看.  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.

视频: 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).

增强容器成像MicroCT

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

科研

医学

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.

实时成像在乳腺癌小鼠模型中的肿瘤微环境对药物的反应

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.

在活体小鼠的血管炎症和血栓形成的异型血小板中性粒细胞被激活的内皮细胞相互作用对实时成像

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

免疫与感染

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.

在肿瘤血管生成中使用荧光共聚焦电视显微镜活体成像

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.

一种新型的高分辨率<em&gt;在体内</em&gt;成像技术研究肿瘤生长及治疗颅内结构的动态响应

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

制备基于微流体的血管化人类微肿瘤模型

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.

作者聚焦：创建人类血管化微肿瘤作为转化癌症研究的模型 

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.

建立用于癌症研究的生理性人血管化微肿瘤模型

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

癌症研究

小鼠的引导肿瘤接种和多模式成像

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.

作者聚焦:整合高分辨率活体成像和 MRI 以增强立体定向放射治疗计划

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.

背侧皮褶窗室肿瘤小鼠模型在转化癌症研究中用于联合活体显微镜和磁共振成像

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

化疗引起的血管毒性 - 实时

摘要

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增强容器成像MicroCT

实时成像在乳腺癌小鼠模型中的肿瘤微环境对药物的反应

在活体小鼠的血管炎症和血栓形成的异型血小板中性粒细胞被激活的内皮细胞相互作用对实时成像

在肿瘤血管生成中使用荧光共聚焦电视显微镜活体成像

一种新型的高分辨率

建立用于癌症研究的生理性人血管化微肿瘤模型

建立用于癌症研究的生理性人血管化微肿瘤模型

建立用于癌症研究的生理性人血管化微肿瘤模型

背侧皮褶窗室肿瘤小鼠模型在转化癌症研究中用于联合活体显微镜和磁共振成像

背侧皮褶窗室肿瘤小鼠模型在转化癌症研究中用于联合活体显微镜和磁共振成像