Name: acellular and cellular lung model to study tumor metastasis
Uploaded: 2018-08-19T14:00:00.000+00:00
Duration: 8 min 31 s
Description: Watch this Scientific Journal Video about Acellular and Cellular Lung Model to Study Tumor Metastasis at JoVE.com

كيم ص دقيقة تلقي الدعم منحة من "جون الثاني جورج منحة البحث كيركلين"، الرابطة الأمريكية لمنح الصدر جراحة، غراهام مؤسسة، هيوستن الميثودية تخصص الطبيب فريق الأبحاث، ومايكل م. وجوان H. مخروط جائزة البحث. ونحن نشكر سيكين Ann لتحرير اللغة من المخطوطة.

<ol>
	<li>Siegel, R. L., Miller, K. D., Jemal, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+Statistics,+2017.">Cancer Statistics, 2017.</a> CA: A Cancer Journal for Clinicians. 67 (1), 7-30 (2017).</li><li>Torre, L. A., Siegel, R. L., Jemal, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lung+Cancer+Statistics.">Lung Cancer Statistics.</a> Advances in Experimental Medicine and Biology. 893, 1-19 (2016).</li><li>Lallo, A., Schenk, M. W., Frese, K. K., Blackhall, F., Dive, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Circulating+tumor+cells+and+CDX+models+as+a+tool+for+preclinical+drug+development.">Circulating tumor cells and CDX models as a tool for preclinical drug development.</a> Translational Lung Cancer Research. 6 (4), 397-408 (2017).</li><li>Yang, S., Zhang, J. J., Huang, X. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+for+tumor+metastasis.">Mouse models for tumor metastasis.</a> Methods in Molecular Biology. 928, 221-228 (2012).</li><li>Bissell, M. J., Hines, W. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Why+don't+we+get+more+cancer?+A+proposed+role+of+the+microenvironment+in+restraining+cancer+progression.">Why don't we get more cancer? A proposed role of the microenvironment in restraining cancer progression.</a> Nature Medicine. 17 (3), 320-329 (2011).</li><li>Francia, G., Cruz-Munoz, W., Man, S., Xu, P., Kerbel, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+advanced+spontaneous+metastasis+for+experimental+therapeutics.">Mouse models of advanced spontaneous metastasis for experimental therapeutics.</a> Nature Reviews Cancer. 11 (2), 135-141 (2011).</li><li>Mishra, D. K., 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=Human+lung+cancer+cells+grown+in+an+ex+vivo+3D+lung+model+produce+matrix+metalloproteinases+not+produced+in+2D+culture.">Human lung cancer cells grown in an ex vivo 3D lung model produce matrix metalloproteinases not produced in 2D culture.</a> PloS One. 7 (9), e45308 (2012).</li><li>Vishnoi, M., Mishra, D. K., Thrall, M. J., Kurie, J. M., Kim, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Circulating+tumor+cells+from+a+4-dimensional+lung+cancer+model+are+resistant+to+cisplatin.">Circulating tumor cells from a 4-dimensional lung cancer model are resistant to cisplatin.</a> The Journal of Thoracic and Cardiovascular Surgery. 148 (3), 1056-1063 (2014).</li><li>Mishra, D. K., 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=Gene+expression+profile+of+A549+cells+from+tissue+of+4D+model+predicts+poor+prognosis+in+lung+cancer+patients.">Gene expression profile of A549 cells from tissue of 4D model predicts poor prognosis in lung cancer patients.</a> International Journal of Cancer. Journal International du Cancer. , (2013).</li><li>Mishra, D. K., 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=Ex+vivo+four-dimensional+lung+cancer+model+mimics+metastasis.">Ex vivo four-dimensional lung cancer model mimics metastasis.</a> The Annals of Thoracic Surgery. 99 (4), 1149-1156 (2015).</li><li>Mishra, D. K., 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=Human+lung+cancer+cells+grown+on+acellular+rat+lung+matrix+create+perfusable+tumor+nodules.">Human lung cancer cells grown on acellular rat lung matrix create perfusable tumor nodules.</a> The Annals of Thoracic Surgery. 93 (4), 1075-1081 (2012).</li><li>Pence, K. A., Mishra, D. K., Thrall, M., Dave, B., Kim, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Breast+cancer+cells+form+primary+tumors+on+ex+vivo+four-dimensional+lung+model.">Breast cancer cells form primary tumors on ex vivo four-dimensional lung model.</a> Journal of Surgical Research. 210, 181-187 (2017).</li><li>Mishra, D. K., 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=Human+Lung+Fibroblasts+Inhibit+Non-Small+Cell+Lung+Cancer+Metastasis+in+Ex+Vivo+4D+Model.">Human Lung Fibroblasts Inhibit Non-Small Cell Lung Cancer Metastasis in Ex Vivo 4D Model.</a> The Annals of Thoracic Surgery. 100 (4), 1167-1174 (2015).</li><li>Mishra, D. K., Miller, R. A., Pence, K. A., Kim, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small+cell+and+non+small+cell+lung+cancer+form+metastasis+on+cellular+4D+lung+model.">Small cell and non small cell lung cancer form metastasis on cellular 4D lung model.</a> BMC Cancer. 18 (1), 441 (2018).</li></ol>

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الرئة 4-د السابقين فيفو يوفر فرصة لدراسة نمو الورم والانبثاث في إنشاء مختبر. مصفوفة الرئة أصلي هو نظام معقد التي تقدم الدعم لانسجة طبيعية ويحافظ على التفاعلات خلية خلية، والخلية-مصفوفة التفاعلات، والتمايز الخلوي وتنظيم الأنسجة. أنه يوفر فرصة لإضافة أي مكونات ورم المكروية لدراسة آثار...

سرطان خبيث هو السبب وراء معظم الوفيات الناجمة عن السرطان، ويشكل التحدي النهائي في الجهود المبذولة لمكافحة السرطان. والهدف العام لهذا الأسلوب هو تصميم بروتوكول لثقافة رباعية الأبعاد (4-د) خلية التي بعدا من التدفق، بالإضافة إلى نمو الخلية (ثلاثي الأبعاد) ثلاثية الأبعاد. وهي تمثل ثلاث مراحل متميزة لعملية ورم خبيث [أي، الورم الرئيسي، تعميم الخلايا السرطانية (ريادية)، وآفات المنتشر].
على مدى العقود الثلاثة الماضية، حقق العلماء في جميع أنحاء العالم ثروة لم يسبق لها مثيل من المعلومات فهم الآليات الكامنة وراء تطور المنتشر في مختلف أنواع السرطان التي تحسن إمكانية علاج أو البقاء خالية من التقدم. الإدارة السريرية لبعض أنواع السرطان، مثل سرطان الثدي، تحسنت بدرجة كبيرة1؛ ومع ذلك، لا تزال لديها بعض أنواع السرطان، مثل سرطان الرئة، وبقاء فقراء2. نماذج حيوانية في المختبر و في فيفو كان لها دور أساسي في توليد رؤى جديدة في الآليات التي تدعم تطوير المرض. في السنوات القليلة الماضية، خلية تكثيفها المستمدة من خط (CDX) وقد تم تكثيفها المستمدة من المريض (PDX) أكثر فائدة كما أنها تحافظ على العديد من الميزات ذات الصلة الورم البشري الرئيسي3، مثل حركية النمو، ميزات النسيجي، والسلوكية الخصائص، والاستجابة للعلاج. ومع ذلك، كل نموذج لها حدودها لفهم إليه تشكيل لجنة مكافحة الإرهاب وورم خبيث إلى جهاز بعيد4،،من56.
في الآونة الأخيرة، قمنا بتطوير نموذج سرطان الرئة 4-د السابقين فيفو باستخدام مفهوم إعادة هندسة الجهاز وثقافة الخلية على أساس التروية. فإنه يحاكي نمو سرطان الرئة البشرية عن طريق تشكيل الورم بيرفوسابل العقيدات التي تنمو على مر الزمن مع إنسان مماثلة السرطان يفرز البروتين إنتاج7. وهو يمثل التوقيع التعبير الجيني الذي تتوقع بقاء الفقراء في المرضى الذين يعانون من السرطان، وكما يبين استجابة علاجية بورم في الانحدار عند سيسبلاتين معاملة8،9. كذلك تعديل نموذج الرئة حيث أن أنها يمكن أن تشكل آفات المنتشر. تتطور من الورم الرئيسي وإينترافاساتي إلى المفرج ريادية واكسترافاساتي في الرئة كونترالاتيرال تشكل آفات المنتشر10. دراسات التعبير الجيني تشير إلى ملف تعريف تعبير متميزة من الورم الرئيسي وريادية، والآفات المنتشر، و upregulation من مجموعة فرعية الجينات المطلوبة ل النمط الظاهري10. تحدث هذه العملية المنتشر بسبب وجود الظروف البيولوجية ينظر في المرضى الذين يعانون من السرطان. وميزة هذا النموذج هو وجود مصفوفة طبيعية والهندسة المعمارية، ونضح المواد الغذائية الذي يؤدي إلى تشكيل الورم العقيدات. وباﻹضافة إلى ذلك، فإنه يوفر أيضا فرصة لدراسة آثار مختلف مكونات وورم المكروية أو المخدرات على تطور الورم على مر الزمن. يمكن استخدام هذا النموذج لنمو مجموعة من الخلايا السرطانية (سرطان الرئة، وسرطان الثدي، ساركومه إلخ) في إنشاء مختبر.

<table><tbody><tr><td>Sprague Dowley rat</td><td>Harlan</td><td>206M</td><td>Male</td></tr><tr><td>Chlorhexidine swab</td><td>Prevantics, NY, USA</td><td>NDC 10819-1080-1</td><td></td></tr><tr><td>Heparin</td><td>Sagent Pharmaceuticals, Schaumburg, IL, USA</td><td>NDC 25021-400-10</td><td></td></tr><tr><td>18-gauge needle</td><td>McMaster Carr, USA</td><td>75165A249</td><td></td></tr><tr><td>2-0 silk tie</td><td>Ethicon, San Angelo, TX, USA</td><td>A305H</td><td></td></tr><tr><td>Masterflex L/S pump</td><td>Cole-Parmer, Vernon Hills, IL, USA</td><td>EW-07554-80</td><td></td></tr><tr><td>Masterflex L/S pump head</td><td>Cole-Parmer, Vernon Hills, IL, USA</td><td>EW-07519-05</td><td></td></tr><tr><td>Masterflex L/S pump cartridge</td><td>Cole-Parmer, Vernon Hills, IL, USA</td><td>EW-07519-70</td><td></td></tr><tr><td>Tygon Tube</td><td>Cole-Parmer, Vernon Hills, IL, USA</td><td>14171211</td><td></td></tr><tr><td>MasterFlex Pump tube</td><td>Cole-Parmer, Vernon Hills, IL, USA</td><td>06598-16</td><td></td></tr><tr><td>Female luer lock connectors</td><td>Cole-Parmer, Vernon Hills, IL, USA</td><td>45508-34</td><td>75165A249</td></tr><tr><td>Male luer lock connectors</td><td>Cole-Parmer, Vernon Hills, IL, USA</td><td>45513-04</td><td></td></tr><tr><td>black nylon ring</td><td>Cole-Parmer, Vernon Hills, IL, USA</td><td>EW-45509-04</td><td></td></tr><tr><td>Intravenous set</td><td>CareFusion</td><td>41134E</td><td></td></tr><tr><td>Sodium Dodecyl Sulfate (SDS)</td><td>Fisher Scientific</td><td>CAS151-21-3</td><td></td></tr><tr><td>Triton X-100</td><td>Sigma-Aldrich</td><td>X100-1L</td><td></td></tr><tr><td>Antibiotics</td><td>Gibco</td><td>15240-062</td><td></td></tr><tr><td>Silicone oxygenator</td><td>Cole-Parmer, Vernon Hills, IL, USA</td><td>ABW00011</td><td>Saint-GoBain-</td></tr><tr><td>Wire mesh</td><td>1164610105</td><td>Lowes</td><td>New York Wire</td></tr><tr><td>Female luer Lug Style TEE</td><td>Cole-Parmer, Vernon Hills, IL, USA</td><td>45508-56</td><td></td></tr><tr><td>Male luer integral lock ring to 200series Barb</td><td>Cole-Parmer, Vernon Hills, IL, USA</td><td>45518-08</td><td></td></tr><tr><td>Female luer thread style coupler</td><td>Cole-Parmer, Vernon Hills, IL, USA</td><td>45508-22</td><td></td></tr><tr><td>Clave connector</td><td>ICU Medical</td><td>11956</td><td></td></tr><tr><td>Hi-Flo &amp;trade;4-way Stopcock w/swivel male luer lock</td><td>smith Medical</td><td>MX9341L</td><td></td></tr><tr><td>MasterFlex Pump tube</td><td>Cole-Parmer, Vernon Hills, IL, USA</td><td>06598-13</td><td>for cannula</td></tr></tbody></table>

acellular and cellular lung model to study tumor metastasis

من الصعب عزل الخلايا السرطانية في مراحل مختلفة من تطور الورم. قمنا بإنشاء نموذج الرئة السابقين فيفو يمكنه إظهار التفاعل بين الخلايا السرطانية مع مصفوفة الطبيعي والتدفق المستمر للمواد الغذائية، وكذلك نموذج يوضح التفاعل بين الخلايا السرطانية مع المكونات الخلوية العادية ومصفوفة طبيعية. يتم إنشاء نموذج الرئة اللاخلوي السابقين فيفو بعزل كتلة قلب والرئة في الفئران وإزالة كافة الخلايا باستخدام عملية ديسيلولاريزيشن. يتم ربط القصبات الرئيسية الحق قبالة وتوضع الخلايا السرطانية في القصبة الهوائية بحقنه. نقل الخلايا وملء الرئة اليسرى. ثم يتم وضع الرئة في مفاعل حيوي حيث يتلقى الشريان الرئوي إلى تدفق مستمر لوسائل الإعلام في دائرة مغلقة. الورم في الرئة اليسرى من الورم الرئيسي. ورم الخلايا التي يتم عزل في وسائط الإعلام يتناقل تنتشر الخلايا السرطانية والخلايا السرطانية في الرئة اليمنى آفات المنتشر. يتم إنشاء نموذج الرئة الخلوية السابقين فيفو بتخطي عملية ديسيلولاريزيشن. يمكن استخدام كل نموذج للإجابة على الأسئلة البحثية المختلفة.

وتحتفظ الرئة حصادها من الفئران سليمة المفرج والحويصلات الهوائية11 (الشكل 3 ألف و 3 باء). عند ديسيلولاريزيشن، يتم الاحتفاظ بعناصر المصفوفة خارج الخلية الرئة اللاخلوي، مثل الكولاجين، فيبرونيكتين، والايلاستين،11 (<strong class="xf...

Watch this Scientific Journal Video about Acellular and Cellular Lung Model to Study Tumor Metastasis at JoVE.com

Acellular and Cellular Lung Model to Study Tumor Metastasis

نموذج الرئة اللاخلوي والخلوية لدراسة ورم خبيث

، نقدم هنا بروتوكولا السابقين فيفو الرئة سرطان نموذج يحاكي خطوات تطور الورم ويساعد على عزل ورم الابتدائي، تعميم خلايا الورم، وآفات المنتشر.

It is difficult to isolate tumor cells at different points of tumor progression. We created an ex vivo lung model that can show the interaction of tumor ...

acellular-and-cellular-lung-model-to-study-tumor-metastasis

Universidad San Sebastian

Research

JoVE Journal

Cancer Research

7.5K Views.  Houston Methodist Hospital Research Institute. ، نقدم هنا بروتوكولا السابقين فيفو الرئة سرطان نموذج يحاكي خطوات تطور الورم ويساعد على عزل ورم الابتدائي، تعميم خلايا الورم، وآفات المنتشر.

Video: Acellular and Cellular Lung Model to Study Tumor Metastasis

Lung Tumor Cell Recruitment Assay

To investigate the molecular mechanisms governing tumor metastasis, various assays using the mouse as a model animal have been proposed. Here, we demonstrate a simple assay to evaluate tumor cell extravasation or micrometastasis. In this assay, tumor cells were injected through the tail vein, and after a short period, the lungs were dissected and digested to count the accumulated labeled tumor cells. This assay skips the initial step of primary tumor invasion into the blood vessel and facilitates the study of events in the distant organ where tumor metastasis occurs. The number of cells injected into the blood vessel can be optimized to observe a limited number of metastases. It has been reported that stromal cells in the distant organ contribute to metastasis. Thus, this assay could be a useful tool to explore potential therapeutic drugs or devices for prevention of tumor metastasis.

This manuscript describes a method to quantify tumor cell accumulation in the lungs in an animal model of tumor metastasis.

الرئة ورم الخلية تجنيد الإنزيم

To investigate the molecular mechanisms governing tumor metastasis, various assays using the mouse as a model animal have been proposed. Here, we ...

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.

المتقدم نموذج حيواني من القولون والمستقيم ورم خبيث في الكبد: تقنيات التصوير وخصائص النقيلي النسخ

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

Experimental Metastasis and CTL Adoptive Transfer Immunotherapy Mouse Model

Experimental metastasis mouse model is a simple and yet physiologically relevant metastasis model. The tumor cells are injected intravenously (i.v) into mouse tail veins and colonize in the lungs, thereby, resembling the last steps of tumor cell spontaneous metastasis: survival in the circulation, extravasation and colonization in the distal organs. From a therapeutic point of view, the experimental metastasis model is the simplest and ideal model since the target of therapies is often the end point of metastasis: established metastatic tumor in the distal organ. In this model, tumor cells are injected i.v into mouse tail veins and allowed to colonize and grow in the lungs. Tumor-specific CTLs are then injected i.v into the metastases-bearing mouse. The number and size of the lung metastases can be controlled by the number of tumor cells to be injected and the time of tumor growth. Therefore, various stages of metastasis, from minimal metastasis to extensive metastasis, can be modeled. Lung metastases are analyzed by inflation with ink, thus allowing easier visual observation and quantification.

An experimental lung metastasis and CTL immunotherapy mouse model for analysis of tumor cells-T cell interaction in vivo.

الإنبثاث التجريبية وCTL المناعي نقل بالتبني النموذجي ماوس

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

Immunology and Infection

Practical Considerations in Studying Metastatic Lung Colonization in Osteosarcoma Using the Pulmonary Metastasis Assay

The pulmonary metastasis assay (PuMA) is an ex vivo lung explant and closed cell culture system that permits researchers to study the biology of lung colonization in osteosarcoma (OS) by fluorescence microscopy. This article provides a detailed description of the protocol, and discusses examples of obtaining image data on metastatic growth using widefield or confocal fluorescence microscopy platforms. The flexibility of the PuMA model permits researchers to study not only the growth of OS cells in the lung microenvironment, but also to assess the effects of anti-metastatic therapeutics over time. Confocal microscopy allows for unprecedented, high-resolution imaging of OS cell interactions with the lung parenchyma. Moreover, when the PuMA model is combined with fluorescent dyes or fluorescent protein genetic reporters, researchers can study the lung microenvironment, cellular and subcellular structures, gene function, and promoter activity in metastatic OS cells. The PuMA model provides a new tool for osteosarcoma researchers to discover new metastasis biology and assess the activity of novel anti-metastatic, targeted therapies.

The goal of this article is to provide a detailed description of the protocol for the pulmonary metastasis assay (PuMA). This model permits researchers to study metastatic osteosarcoma (OS) cell growth in lung tissue using a widefield fluorescence or confocal laser-scanning microscope.

الاعتبارات العملية في دراسة الاستعمار الرئة المنتشر في أوستيوساركوما باستخدام مقايسة خبيث الرئوية

The pulmonary metastasis assay (PuMA) is an ex vivo lung explant and closed cell culture system that permits researchers to study the biology of lung ...

The Establishment of a Lung Colonization Assay for Circulating Tumor Cell Visualization in Lung Tissues

Metastasis is the major cause of cancer death. The role of circulating tumor cells (CTCs) in promoting cancer metastasis, in which lung colonization by CTCs critically contributes to early lung metastatic processes, has been vigorously investigated. As such, animal models are the only approach that captures the full systemic process of metastasis. Given that problems occur in previous experimental designs for examining the contributions of CTCs to blood vessel extravasation, we established an in vivo lung colonization assay in which a long-term-fluorescence cell-tracer, carboxyfluorescein succinimidyl ester (CFSE), was used to label suspended tumor cells and lung perfusion was performed to clear non-specifically trapped CTCs prior to lung removal, confocal imaging, and quantification. Polymeric fibronectin (polyFN) assembled on CTC surfaces has been found to mediate lung colonization in the final establishment of metastatic tumor tissues. Here, to specifically test the requirement of polyFN assembly on CTCs for lung colonization and extravasation, we performed short term lung colonization assays in which suspended Lewis lung carcinoma cells (LLCs) stably expressing FN-shRNA (shFN) or scramble-shRNA (shScr) and pre-labeled with 20 &#956;M of CFSE were intravenously inoculated into C57BL/6 mice. We successfully demonstrated that the abilities of shFN LLC cells to colonize the mouse lungs were significantly diminished in comparison to shScr LLC cells. Therefore, this short-term methodology may be widely applied to specifically demonstrate the ability of CTCs within the circulation to colonize the lungs.

An animal model is needed to decipher the role of circulating tumor cells (CTCs) in promoting lung colonization during cancer metastasis. Here, we established and successfully performed an in vivo assay to specifically test the requirement of polymeric fibronectin (polyFN) assembly on CTCs for lung colonization.

إنشاء مقايسة الاستعمار الرئة لتعميم التصور خلية الورم في أنسجة الرئة

Metastasis is the major cause of cancer death. The role of circulating tumor cells (CTCs) in promoting cancer metastasis, in which lung colonization by ...

Pathological Analysis of Lung Metastasis Following Lateral Tail-Vein Injection of Tumor Cells

Metastasis, the primary cause of morbidity and mortality for most cancer patients, can be challenging to model preclinically in mice. Few spontaneous metastasis models are available. Thus, the experimental metastasis model involving tail-vein injection of suitable cell lines is a mainstay of metastasis research. When cancer cells are injected into the lateral tail-vein, the lung is their preferred site of colonization. A potential limitation of this technique is the accurate quantification of the metastatic lung tumor burden. While some investigators count macrometastases of a pre-defined size and/or include micrometastases following sectioning of tissue, others determine the area of metastatic lesions relative to normal tissue area. Both of these quantification methods can be exceedingly difficult when the metastatic burden is high. Herein, we demonstrate an intravenous injection model of lung metastasis followed by an advanced method for quantifying metastatic tumor burden using image analysis software. This process allows for investigation of multiple end-point parameters, including average metastasis size, total number of metastases, and total metastasis area, to provide a comprehensive analysis. Furthermore, this method has been reviewed by a veterinary pathologist board-certified by the American College of Veterinary Pathologists (SEK) to ensure accuracy.

Intravenous injection of cancer cells is often used in metastasis research, but the metastatic tumor burden can be difficult to analyze. Herein, we demonstrate a tail-vein injection model of metastasis and include a novel approach to analyze the resulting metastatic lung tumor burden.

التحليل المرضي لنقائل الرئة بعد الحقن الجانبي الوريد الذيل من الخلايا السرطانية

Metastasis, the primary cause of morbidity and mortality for most cancer patients, can be challenging to model preclinically in mice. Few spontaneous ...

A Permanent Window for Investigating Cancer Metastasis to the Lung

Metastasis, accounting for ~90% of cancer-related mortality, involves the systemic spread of cancer cells from primary tumors to secondary sites such as the bone, brain, and lung. Although extensively studied, the mechanistic details of this process remain poorly understood. While common imaging modalities, including computed tomography (CT), positron emission tomography (PET), and magnetic resonance imaging (MRI), offer varying degrees of gross visualization, each lacks the temporal and spatial resolution necessary to detect the dynamics of individual tumor cells. To address this, numerous techniques have been described for intravital imaging of common metastatic sites. Of these sites, the lung has proven especially challenging to access for intravital imaging owing to its delicacy and critical role in sustaining life. Although several approaches have previously been described for single-cell intravital imaging of the intact lung, all involve highly invasive and terminal procedures, limiting the maximum possible imaging duration to 6-12 h. Described here is an improved technique for the permanent implantation of a minimally invasive thoracic optical Window for High-Resolution Imaging of the Lung (WHRIL). Combined with an adapted approach to microcartography, the innovative optical window facilitates serial intravital imaging of the intact lung at single-cell resolution across multiple imaging sessions and spanning multiple weeks. Given the unprecedented duration of time over which imaging data can be gathered, the WHRIL can facilitate the accelerated discovery of the dynamic mechanisms underlying metastatic progression and numerous additional biologic processes within the lung.

Here, we present a protocol for the surgical implantation of a permanently indwelling optical window for the murine thorax, which enables high-resolution, intravital imaging of the lung. The permanence of the window makes it well-suited to the study of dynamic cellular processes in the lung, especially those that are slowly evolving, such as metastatic progression of disseminated tumor cells.

نافذة دائمة للتحقيق في انتشار السرطان إلى الرئة

Metastasis, accounting for ~90% of cancer-related mortality, involves the systemic spread of cancer cells from primary tumors to secondary sites such as ...

فحص انتشار الخلايا السرطانية من نقائل الرئة

Tracking Tumor Cell Dissemination from Lung Metastases Using Photoconversion

Metastasis - the systemic spread of cancer - is the leading cause of cancer-related deaths. Although metastasis is commonly thought of as a unidirectional process wherein cells from the primary tumor disseminate and seed metastases, tumor cells in existing metastases can also redisseminate and give rise to new lesions in tertiary sites in a process known as &#34;metastasis-from-metastases&#34; or &#34;metastasis-to-metastasis seeding.&#34; Metastasis-to-metastasis seeding may increase the metastatic burden and decrease the patient&#39;s quality of life and survival. Therefore, understanding the processes behind this phenomenon is crucial to refining treatment strategies for patients with metastatic cancer.
Little is known about metastasis-to-metastasis seeding, due in part to logistical and technological limitations. Studies on metastasis-to-metastasis seeding rely primarily on sequencing methods, which may not be practical for researchers studying the exact timing of metastasis-to-metastasis seeding events or what promotes or prevents them. This highlights the lack of methodologies that facilitate the study of metastasis-to-metastasis seeding. To address this, we have developed - and describe herein - a murine surgical protocol for the selective photoconversion of lung metastases, allowing specific marking and fate tracking of tumor cells redisseminating from the lung to tertiary sites. To our knowledge, this is the only method for studying tumor cell redissemination and metastasis-to-metastasis seeding from the lungs that does not require genomic analysis.

تسليط الضوء على المؤلف: فك تشفير البذر من ورم خبيث إلى ورم خبيث باستخدام تقنية جديدة في الجسم الحي لتتبع انتشار سرطان الثدي 

We present a method for studying tumor cell redissemination from lung metastases involving a surgical protocol for the selective photoconversion of lung metastases, followed by the identification of redisseminated tumor cells in tertiary organs.

تتبع انتشار الخلايا السرطانية من نقائل الرئة باستخدام التحويل الضوئي

Metastasis - the systemic spread of cancer - is the leading cause of cancer-related deaths. Although metastasis is commonly thought of as a unidirectional ...

نموذج الرئة اللاخلوي والخلوية لدراسة ورم خبيث

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الرئة ورم الخلية تجنيد الإنزيم

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الاعتبارات العملية في دراسة الاستعمار الرئة المنتشر في أوستيوساركوما باستخدام مقايسة خبيث الرئوية

إنشاء مقايسة الاستعمار الرئة لتعميم التصور خلية الورم في أنسجة الرئة

التحليل المرضي لنقائل الرئة بعد الحقن الجانبي الوريد الذيل من الخلايا السرطانية

نافذة دائمة للتحقيق في انتشار السرطان إلى الرئة

تتبع انتشار الخلايا السرطانية من نقائل الرئة باستخدام التحويل الضوئي

تتبع انتشار الخلايا السرطانية من نقائل الرئة باستخدام التحويل الضوئي

تتبع انتشار الخلايا السرطانية من نقائل الرئة باستخدام التحويل الضوئي