Name: practical considerations in studying metastatic lung colonization in osteosarcoma using the pulmonary metastasis assay
Uploaded: 2018-03-12T13:00:00
Description: Watch this Scientific Journal Video about Practical Considerations in Studying Metastatic Lung Colonization in Osteosarcoma Using the Pulmonary Metastasis Assay at JoVE.com

Wir möchten danken Dr. Arnulfo Mendoza in der PuMA-Technik geschult. Darüber hinaus möchten wir DRS. Chand Khanna, Susan Garfield (NCI/NIH) und Sam Aparicio (BC Cancer Agency) für die Nutzung von ihren Mikroskopen im Laufe dieser Studie anerkennen. Diese Forschung wurde von Intramural Research Program des National Institutes of Health, Center for Cancer Research, Pädiatrische Onkologie-Zweig (teilweise) unterstützt. M.M.L. vom nationalen Instituten der Gesundheit intramuralen Besuch Fellow Programm (Award 15335) unterstützt wurde und wird derzeit von einem Joan-Parker-Stipendium in Metastasen Forschung unterstützt. P.H.S. wird von der British Columbia Cancer Foundation unterstützt.

Alle tierischen Protokolle aus denen Bilddaten stammen wurden mit Genehmigung der Animal Care und Use Committee des National Cancer Institute, National Institutes of Health durchgeführt. Alle tierischen Protokolle diskutiert und dargestellt in dem Artikel video wurden von der University of British Columbia Animal Care Ausschuss genehmigt.
1. Vorbereitung der Tumorzellen für Einspritzung und Materialien für das PuMA-Modell
Hinweis: Die Menge der Lösungen und Zellen...

<ol>
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Die folgende technischen Artikel beschreibt einige praktische Aspekte des Modells bei der Untersuchung der Lunge Kolonisation in OS PuMA. Im folgenden: einige wichtige Schritte in das Protokoll, wo Forscher besonders vorsichtig nehmen sollte
(a) Kanülierung der Luftröhre. Die Luftröhre kann leicht beschädigt werden, während sie sezieren die umliegenden Muskeln und Bindegewebe. Darüber hinaus kann die Nadel des Katheters leicht durch die Luftröhre gedrückt werden. Achten Sie besonders a...

Bessere Ergebnisse für pädiatrische Patienten mit metastasierendem Osteosarkom (OS) bleibt noch ein kritischer ungedeckten klinischen Bedarf 1. Dies unterstreicht die Bedeutung der Entwicklung neuer molekular zielgerichtete Therapien. Herkömmliche Chemotherapeutika, die Ziel-Tumor-Zell-Proliferation haben nicht als wirksam erwiesen bei der Behandlung von Metastasen und damit neue Strategien muss der metastatischen Prozess selbst 2ausgerichtet. Der aktuelle Artikel beschreibt die praktischen Aspekte der eine relativ neue Art von ex Vivo Lunge Metastasen Modell, entwickelt von Mendoza und Kollegen3, wonach ein nützliches Werkzeug bei der Entdeckung neuer pulmonalen Metastasen-Assay (PuMA) molekularen Treiber in der Lunge Metastasen Progression in OS 4,5. Bevor Sie fortfahren, es wäre klug, kurz auf einige aktuelle Modelle von Metastasen zu berühren, und wie das PuMA-Modell bietet mehrere Vorteile gegenüber konventionellen in-vitro- Tests.
Die meisten experimentelle Modellen genutzt, um Metastasen zu untersuchen umfassen in Vitro und in Vivo -Systeme, die entweder einem bestimmten Schritt oder mehrere Stufen der metastatischen Kaskade zu rekapitulieren. Diese Schritte umfassen: (1) die Migration vom Primärtumor, 2) Intravasation in nahe gelegenen Gefäße (Blut- oder Lymphgefäße) und Transitländern im Umlauf, Tumorzellen (3) verhaften am sekundären Standort, 4) Extravasation und Überleben am sekundären Standort, Bildung (5) Fixierungsprotokoll, und 6) Wachstum in vaskularisierte Metastasen (Abbildung 1). In-vitro- Modelle von Metastasen können 2-dimensionale (2D) Migration und 3-dimensionale (3D) Matrigel Invasion Assays, die in überprüft werden ausführlich an anderer Stelle 6. Für in-Vivo -Modelle, die zwei häufig verwendete Modellsysteme umfassen: 1) das spontane Metastasierung Modell ist wo ein Tumor-Zellen sind Orthotopically, die in einem bestimmten Gewebetyp einen lokalen Tumor bilden, die spontan metastasierende Zellen wirft injiziert an entfernten Standorten; (2) die experimentelle Metastasierung Modell ist wo Tumorzellen in das Blutgefäß stromaufwärts von dem Zielorgan injiziert werden. Zum Beispiel eine Rute Vene Injektion von Tumor-Zellen-Ergebnisse in der Entwicklung Lunge Metastasen5,7,8. Andere experimentelle Metastasen-Modelle verfügen über Injektion von Tumorzellen in die Milz oder mesenterialen Vene führt zu die Entwicklung von Lebermetastasen9,10. Praktische Erwägungen dieser in-Vivo -Modelle werden von Welch 11ausführlich erörtert. Ein weiteres in-Vivo -Modell zur Untersuchung von Metastasen in den pädiatrischen Sarkome ist der Nieren Nieren subkapsuläre Tumor Implantation Modell ergibt sich lokale Tumorbildung und spontane Metastasen in der Lunge 12,13. Eine technisch anspruchsvolle Technik wie intravitalen Videomicroscopy können direkt sichtbar zu machen, in Echtzeit, Wechselwirkungen zwischen metastatischen Krebszellen und den Microvasculature einer metastatischen Website (dh. Lunge oder Leber) wie beschrieben von MacDonald14 und Entenberg15oder Krebs Zelle Extravasation in der chorioallantoic Membrane wie beschrieben durch Kim 16.
Das PuMA-Modell ist ein ex-Vivo, Lunge Gewebe Explant, geschlossenes Kultursystem wo kann das Wachstum von Tumorzellen fluoreszierende längs über Fluoreszenz-Mikroskopie über einen Zeitraum von einem Monat beobachtet werden (siehe Abb. 2A). Dieses Modell rekapituliert die Anfangsphase der Lunge Kolonisation (Schritte 3-5) in der metastasierten Kaskade. Einige große Vorteile gegenüber konventionellen in-vitro- Modellen des Modells PuMA sind: 1) Es bietet die Möglichkeit, metastasiertem Krebs Zelle Wachstum in einem 3D Mikroumgebung längs zu messen, die viele Funktionen der Lunge Mikroumgebung behält Vivo 3; (2) PuMA ermöglicht den Forscher zu beurteilen, ob der Zuschlag einer Kandidaten-gen oder Drogenmissbrauch Behandlung Anti-metastatischen Aktivitäten im Zusammenhang mit einem 3D Lunge Mikroumgebung hat; (3) das PuMA-Modell sind flexibel mit vielen Arten von Fluoreszenz-Mikroskopie-Plattformen (Abbildung 2 b) wie Weitfeld-Fluoreszenz-Mikroskopie oder konfokale Laser-scanning-Mikroskopie, Beispiele für jede Abbildung 2 & Dzeigt, bzw.. Dieser Artikel wird erläutert, wie mithilfe der PuMA-Modell um längs Bilddaten auf metastasiertem Wachstum der erweiterten grün fluoreszierendes Protein (eGFP) zu erhalten-mit dem Ausdruck, menschliche Ebbe und metastasierendem Osteosarkom Zellen (MNNG und HOS Zellen) mit niedrig-Vergrößerung Weitfeld Fluoreszenz. Beispiele für imaging ein Fluoreszenzfarbstoff, der Lungenparenchyms Etiketten und ein rot-fluoreszierende Protein, die genetische Reporter der Mitochondrien in OS Etiketten im PuMA Modell konfokale Laser-scanning-Mikroskopie mit Zellen werden ebenfalls besprochen.

<table>
	<tbody>
		<tr>
			<td>Table 2</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cell culture reagents for A-media, B-media, and complete media</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>MNNG-HOS</td>
			<td>ATCC</td>
			<td>CRL-1547</td>
			<td>highly metastatic OS cell line</td>
		</tr>
		<tr>
			<td>HOS</td>
			<td>ATCC</td>
			<td>CRL-1543</td>
			<td>poorly metastatic OS cell line</td>
		</tr>
		<tr>
			<td>MG63.3</td>
			<td>Amy LeBlanc Laboratory (NCI)</td>
			<td>N/A</td>
			<td>highly metastatic OS cell line</td>
		</tr>
		<tr>
			<td>MG63</td>
			<td>ATCC</td>
			<td>CRL-1427</td>
			<td>poorly metastatic OS cell line</td>
		</tr>
		<tr>
			<td>10X M199 media</td>
			<td>Thermofisher</td>
			<td>11825015</td>
			<td>Base media for A-media and B-media</td>
		</tr>
		<tr>
			<td>Distilled Water (sterilized)</td>
			<td>Thermofisher</td>
			<td>15230-147</td>
			<td>Component of A-media &#38; B-media</td>
		</tr>
		<tr>
			<td>7.5% sodium bicarbonate solution</td>
			<td>Thermofisher</td>
			<td>25080094</td>
			<td>Component of A-media &#38; B-media</td>
		</tr>
		<tr>
			<td>Hydrocortizone</td>
			<td>Sigma-Alrich</td>
			<td>H6909</td>
			<td>Component of A-media &#38; B-media</td>
		</tr>
		<tr>
			<td>Retinol acetate-water soluable</td>
			<td>Sigma-Alrich</td>
			<td>R0635-5MG</td>
			<td>Component of A-media &#38; B-media</td>
		</tr>
		<tr>
			<td>Penicillin/Streptomycin 10X concentrated (10000 U/ml) solution</td>
			<td>Thermofisher</td>
			<td>15140122</td>
			<td>Component of A-media &#38; B-media, complete media.</td>
		</tr>
		<tr>
			<td>Bovine insulin solution (10mg/ml)</td>
			<td>Sigma-Alrich</td>
			<td>I0516-5ML</td>
			<td>Component of A-media &#38; B-media</td>
		</tr>
		<tr>
			<td>DMEM, high glucose</td>
			<td>Thermofisher</td>
			<td>11965092</td>
			<td>Base media of Complete Media</td>
		</tr>
		<tr>
			<td>L-Glutamine (200 mM)</td>
			<td>Thermofisher</td>
			<td>25030081</td>
			<td>Component of Complete Media</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum</td>
			<td>Thermofisher</td>
			<td>16000044</td>
			<td>Component of Complete Media</td>
		</tr>
		<tr>
			<td>Dulbecco&#8217;s Phosphate Buffered Saline</td>
			<td>Thermofisher</td>
			<td>14190144</td>
			<td>Used in cell culture.</td>
		</tr>
		<tr>
			<td>Hank&#8217;s Buffered Salts Solution, no calcium, no magnesium, no phenol red</td>
			<td>Thermofisher</td>
			<td>14175095</td>
			<td>Used to resuspend cell pellet prior to injection</td>
		</tr>
		<tr>
			<td>Trypsin-EDTA (0.25%), phenol red</td>
			<td>Thermofisher</td>
			<td>25200114</td>
			<td>Used in cell culture.</td>
		</tr>
		<tr>
			<td>DAR4M</td>
			<td>Enzo</td>
			<td>ALX-620-069-M001</td>
			<td>Used to label lung parenchyma.</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Table 3</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Materials for PuMA</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Zeiss 710 Confocal LSM</td>
			<td>Zeiss</td>
			<td>N/A</td>
			<td>Upright LSM confocal microscope</td>
		</tr>
		<tr>
			<td>Zeiss 780 Confocal LSM</td>
			<td>Zeiss</td>
			<td>N/A</td>
			<td>Inverted LSM confocal microscope</td>
		</tr>
		<tr>
			<td>SCID mice</td>
			<td>Charles River</td>
			<td>N/A</td>
			<td>NOD.CB17-Prkdcscid/NcrCrl, female, age 6-8 weeks</td>
		</tr>
		<tr>
			<td>GelFoam</td>
			<td>Harvard Apparatus</td>
			<td>59-9863</td>
			<td>Used as a support for lung tissue sections.</td>
		</tr>
		<tr>
			<td>SeaPlaque Agarose</td>
			<td>Lonza</td>
			<td>50100</td>
			<td>Used during insufflation of the lung.</td>
		</tr>
		<tr>
			<td>1 ml syringe with 27 gauge needle</td>
			<td>Fisherscientific</td>
			<td>14-826-87</td>
			<td>Used for tail vein injection.</td>
		</tr>
		<tr>
			<td>10 ml syringe</td>
			<td>BD</td>
			<td>309604</td>
			<td>Used for insufflation of the lung.</td>
		</tr>
		<tr>
			<td>20 gauge catheter</td>
			<td>Terumo</td>
			<td>SR-OX2032CA</td>
			<td>Used during insufflation of the lung.</td>
		</tr>
		<tr>
			<td>Abbott IV extension set (30&#34;, Sterile)</td>
			<td>Medisca</td>
			<td>8342</td>
			<td>Used during insufflation of the lung.</td>
		</tr>
		<tr>
			<td>Alcohol swabs</td>
			<td>BD</td>
			<td>326895</td>
			<td>For wiping tail vein before injection</td>
		</tr>
		<tr>
			<td>Sterile surgical gloves</td>
			<td>Fisherscientific</td>
			<td>Varies with size</td>
			<td>Asceptic handing of mouse lungs</td>
		</tr>
		<tr>
			<td>30 cm ruler</td>
			<td>Staples</td>
			<td></td>
			<td>Used for insufflation of the lung.</td>
		</tr>
		<tr>
			<td>Support stand for ruler</td>
			<td>Pipette.com</td>
			<td>HS29022A</td>
			<td>Used for insufflation of the lung.</td>
		</tr>
		<tr>
			<td>35 mm glass-bottomed culture dish</td>
			<td>Ibidi</td>
			<td>81158</td>
			<td>Used during imaging of lung slices</td>
		</tr>
		<tr>
			<td>Absorbent Underpads with Waterproof Moisture Barrier</td>
			<td>VWR</td>
			<td>56617-014</td>
			<td>Used to line the sterile work area in the biological hood.</td>
		</tr>
		<tr>
			<td>Catgut Plain Absorbable Suture</td>
			<td>Braun</td>
			<td>N/A</td>
			<td>Used to tie off cannulated trachea.</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Table 4</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical instruments for PuMA</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Micro Dissecting Scissors 3.5&#34; Straight Sharp/Sharp</td>
			<td>Roboz</td>
			<td>RS-5910</td>
			<td>For cutting lung sections</td>
		</tr>
		<tr>
			<td>4&#8221; (10 cm) Long Serrated Straight Extra Delicate 0.5mm Tip</td>
			<td>Roboz</td>
			<td>RS-5132</td>
			<td>For manipulating/holding lung sections.</td>
		</tr>
		<tr>
			<td>4&#8221; (10 cm) Long Serrated Slight Curve 0.8mm Tip</td>
			<td>Roboz</td>
			<td>RS5135</td>
			<td>For manipulating/holding lung sections.</td>
		</tr>
		<tr>
			<td>Thumb Dressing Forceps; Serrated; Delicate; 4.5&#34; Length; 1.3 mm Tip Width</td>
			<td>Roboz</td>
			<td>RS-8120</td>
			<td>For general dissection.</td>
		</tr>
		<tr>
			<td>Thumb Dressing Forceps 4.5&#34; Serrated 2.2 mm Tip Width</td>
			<td>Roboz</td>
			<td>RS-8100</td>
			<td>For general dissection.</td>
		</tr>
		<tr>
			<td>Extra Fine Micro Dissecting Scissors 3.5&#34; Straight Sharp/Sharp, 20mm blade</td>
			<td>Roboz</td>
			<td>RS-5880</td>
			<td>For general dissection.</td>
		</tr>
		<tr>
			<td>Knapp Scissors; Straight; Sharp-Blunt; 27mm Blade Length; 4&#34; Overall Length</td>
			<td>Roboz</td>
			<td>RS-5960</td>
			<td>For general dissection.</td>
		</tr>
	</tbody>
</table>

practical considerations in studying metastatic lung colonization in osteosarcoma using the pulmonary metastasis assay

Pulmonalen Metastasen-Assay (PuMA) ist ein ex-Vivo Lunge Explant und geschlossenen Zellkultursystem, die es Forschern erlaubt, die Biologie der Lunge Besiedlung in Osteosarkom (OS) durch Fluoreszenz-Mikroskopie zu studieren. Dieser Artikel enthält eine ausführliche Beschreibung des Protokolls und erläutert, Beispiele von Bilddaten auf metastasiertem Wachstum mit Weitfeld oder konfokale Fluoreszenz-Mikroskopie-Plattformen zu erhalten. Die Flexibilität des Modells PuMA erlaubt Forschern, nicht nur das Wachstum von OS-Zellen in der Lunge Mikroumgebung zu studieren, sondern auch die Auswirkungen der anti-metastatischen Therapeutika im Zeitverlauf zu beurteilen. Konfokale Mikroskopie ermöglicht eine noch nie da gewesenen, hochauflösende Bildgebung von OS-Zell-Interaktionen mit dem Lungenparenchym. Darüber hinaus Wenn das PuMA-Modell mit Fluoreszenzfarbstoffen oder fluoreszierende Protein genetische Reporter kombiniert wird, können Forscher die Lunge Mikroumgebung, zellulärer und subzellulärer Strukturen und Genfunktion Promotoraktivität in metastasierenden OS Zellen untersuchen. Das PuMA-Modell bietet ein neues Tool für Osteosarkom-Forscher entdecken neue Metastasen Biologie und die Aktivität der neuartigen anti-metastatischen, gezielte Therapien zu bewerten.

Low-Vergrößerung Weitfeld-Fluoreszenz-Mikroskopie
Für Weitfeld-Fluoreszenz-Mikroskopie von PuMA Lung Slices sind repräsentative Bilder und Quantifizierung Daten in Abbildung 2und Abbildung 4A und Bdargestellt. Die metastatischen Neigungen für hohe und niedrige metastasierendem Zelllinien sind über progressive Zeitpunkten ...

Watch this Scientific Journal Video about Practical Considerations in Studying Metastatic Lung Colonization in Osteosarcoma Using the Pulmonary Metastasis Assay at JoVE.com

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

Das Ziel dieses Artikels ist es, eine detaillierte Beschreibung des Protokolls für die pulmonalen Metastasen-Assay (PuMA) zur Verfügung zu stellen. Dieses Modell erlaubt Forschern, metastasierendem Osteosarkom (OS) Zellwachstum im Lungengewebe mit Weitfeld-Fluoreszenz oder konfokale Laserscanning-Mikroskop zu studieren.

Praktische Überlegungen an einem Studium von metastasierendem Lungenkrebs Besiedlung in Osteosarkom mit pulmonalen Metastasen-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 ...

The overall goal of this pulmonary metastasis assay is to directly visualize and study the growth of metastatic osteosarcoma cells in viable lung tissue. This method can help answer really key questions in the osteosarcoma metastasis field. Such as, how metastatic osteosarcoma cells adapt, survive, and proliferate in the lung micro environment.The main advantage of this technique is that it permits a direct visualization of metastatic osteosarcoma cell growth in a relevant three dimensional micro environment. Generally, individuals new to this method will struggle, because of the technically challenging steps of cannulating the trachea and insufflating the lung. The tail vein injection of tumor cells and mouse euthanasia are done according to standard institutional guidelines.After five minutes, place the euthanized mouse in dorsal recumbency on a sterile pad in a laminar flow hood, and use small, sterile scissors to carefully dissect out the sternum, past the thoracic inlet on both sides of the trachea, without puncturing the lungs. Dissecting all of the muscle and connective tissue surrounding the trachea is essential for a successful cannulation. Next, use a 20 gauge IV catheter to carefully cannulate the trachea and use a sterile catgut suture to loosely secure the catheter and the trachea.Attach an IV extension set from the catheter to the 10 milliliter syringe of a gravity profusion apparatus and pour 37 degrees celsius liquid agarose mixed with A medium solution into the syringe. Fill the lungs with the agarose solution until they are fully insufflated. Then remove the cannula and firmly tie off the suture to prevent leakage of the agarose solution.It is important to keep the lung intact during the insufflation. A proper insufflation with agarose and subsequent cooling of the pluck, will maintain the lung in an expanded state. Harvest the pluck, trachea, heart, and lung, taking care not to puncture or damage the surface of the lung.Place the tissues in 30 milliliters of pre-chilled PBS supplemented with antibiotics, and allow the agarose to solidify. Place the pluck on ice for 20 minutes. After 20 minutes, use fine scissors and tweezers to cut the lung into 3 x 1.5 millimeter pieces, and place the slices into individual wells of a 6 well plate containing 2 x 2 centimeter gelatin sponges pre-soaked in B medium.On the day of imaging, carefully transfer the lung slices from the 6 well plate, into a sterile 35 millimeter glass bottom round dish in columns according to their experimental group. Next, on a wide field fluorescence microscope, select the 2.5x objective and set the imaging parameters to provide the best contrast between the fluorescent tumor cells and the non fluorescing background tissue in the control tissue sample group. Image all of the samples using the same parameters as for the control tissues.If the software isn't scale calibrated, image a micrometer using the same objective as the lung tissue in order to obtain a scale reference, and save all of the images as tif files. When all of the images have been obtained, to analyze the images in ImageJ, open the first file. Use the polygon selection tool to outline the entire lung slice to determine the area of the total lung slice.Select process and subtract background. Set the rolling ball radius to a starting value of 50 pixels. This value will be dependent on the degree of background fluorescence and should be optimized for each data set.Make sure to uncheck the light background. Select image and type, 8-bit, then image and properties, enter pixels. Under unit of length, enter 1 for the pixel width, pixel height, and voxel depth.Then, check global to apply these parameters to all of the subsequent images. Next, select image, then adjust, then threshold. Highlight default and black and white, and use the slider to threshold the image so that the majority of the tumor cells are accurately highlighted.Click apply one time to turn the lung black and the fluorescent lesions white. Then, click apply a second time to invert the image so that all of the fluorescent structures turn black. To quantify the number and shape of the lesions, select analyze and set measurements, and check area.Uncheck all of the other boxes. Then, select analyze particles and in the size field use the area of the smallest lesion determined to be a single cell by ImageJ, to set the range of shapes that the program will enumerate. Use an image from day zero vehicle group to choose the smallest area that is considered to be a single tumor cell.When the lower limit has been set, click okay. A results window will pop up containing all of the enumerated shapes and area measurements. Then, copy and paste the data from the results window into a spreadsheet, and use the sum mathematical function to add all of the areas of the metastatic lesions for that particular lung slice.The metastatic propensities for high and low metastatic cell lines are visually apparent over progressive time points. More specifically, these microscopy images show how highly metastatic cells are able to colonize the lung more efficiently than low metastatic cell lines. Image analysis, quantification, and statistical analysis of the image data confirms that highly metastatic tumor cells can efficiently colonize the lung tissue.Whereas low metastatic cells cannot grow at all. High magnification confocal fluorescence microscopy allows visualization of the lung parenchyma cells in great spatial detail compared to EGFP expressing tumor cells. As well as facilitates the analysis of subcellular structures, such as these mitochondria.Depending on the number of mice used, this technique can be completed in four hours if it is performed properly. While attempting this procedure it is important to remember to work with clean, sterile surgical equipment. After it's development, the technique paved the way for researchers in the field of cancer research to explore metastasis progression in the lungs of mice.Following this procedure, other methods like in vitro cell culture assays and western blotting can be performed to answer additional questions about the effects of gene knockdown or drug treatment on cancer cells. After watching this video, you should have a good understanding of how to perform a pulmonary metastasis assay. Don't forget that working with human cancer cells can be extremely hazardous, and that precautions such as working in a bio level safety two laboratory, should always be taken while performing this procedure.

Research

JoVE Journal

Cancer Research

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

Erweiterte Tiermodell für Colorectal Metastase in der Leber: Imaging-Techniken und Eigenschaften des metastasierten Clones

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

Forschung

Krebsforschung

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

Die Einrichtung eines Lunge Kolonisation Assays für zirkulierenden Tumor Zelle Visualisierung im Lungengewebe

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

Modeling Osteosarcoma Using Li-Fraumeni Syndrome Patient-derived Induced Pluripotent Stem Cells

Modellierung von Osteosarkom mit Li-Fraumeni-Syndrom, die Patienten abgeleitet induzierten pluripotenten Stammzellen

Li-Fraumeni syndrome (LFS) is an autosomal dominant hereditary cancer disorder. Patients with LFS are predisposed to a various type of tumors, including ...

Acellular and Cellular Lung Model to Study Tumor Metastasis

Azelluläre und zellulären Lunge Modell Tumor Metastasen zu studieren

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 ...

Micromanipulation of Circulating Tumor Cells for Downstream Molecular Analysis and Metastatic Potential Assessment

Mikromanipulation von zirkulierenden Tumorzellen für die nachgelagerte Molekulare Analyse und die metastasierende Potenzialbewertung

Blood-borne metastasis accounts for&#160;most cancer-related deaths and involves circulating tumor cells (CTCs) that are successful in establishing new ...

Combined Use of Tail Vein Metastasis Assays and Real-Time In Vivo Imaging to Quantify Breast Cancer Metastatic Colonization and Burden in the Lungs

Kombinierte Verwendung von Tail Vein Metastasis Assays und Real-Time In Vivo Imaging zur Quantifizierung der metastasierenden Kolonisation und Belastung von Brustkrebs in der Lunge

Metastasis is the main cause of cancer-related deaths and there are limited therapeutic options for patients with metastatic disease. The identification ...

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

Pathologische Analyse der Lungenmetastasierung nach lateraler Schwanzveneninjektion von Tumorzellen

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

Using Computer-based Image Analysis to Improve Quantification of Lung Metastasis in the 4T1 Breast Cancer Model

Verwendung computerbasierter Bildanalyse zur Verbesserung der Quantifizierung von Lungenmetastasen im 4T1-Brustkrebsmodell

Breast cancer is a devastating malignancy, accounting for 40,000 female deaths and 30% of new female cancer diagnoses in the United States in 2019 alone. ...

A Syngeneic Orthotopic Osteosarcoma Sprague Dawley Rat Model with Amputation to Control Metastasis Rate

Ein syngenes orthotopes Osteosarkom Sprague Dawley Rattenmodell mit Amputation zur Kontrolle der Metastasenrate

The most recent advance in the treatment of osteosarcoma (OS) occurred in the 1980s when multi-agent chemotherapy was shown to improve overall survival ...

A Permanent Window for Investigating Cancer Metastasis to the Lung

Ein permanentes Fenster zur Untersuchung von Krebsmetastasen in der Lunge

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