Diese Arbeit wurde von der National Science Foundation CAREER-Award (CBET-0846395) und der Abteilung für Vergleichende Medizin, Tulane University gefördert.

1. Animals <ol><li> Weibliche C57BL/6J-Mäuse (Mus musculus) verifiziert werden frei von ansteckenden Ektoparasiten Endoparasiten Helminthen und Antikörper zu 17 murine Viren durch den Verkäufer vor dem Versand. Mäuse sind in der AAALAC-akkreditierten Tierhaltung der Tulane University in Übereinstimmung mit Anleitung für die Pflege und Verwendung von Labortieren untergebracht. 1 </li><li> Genauer gesagt, werden die Mäuse in Gruppen gehalten (4-5 Mäuse / Käfig) in individuell belüfteten Polycarbonat cageswith microisolation Filterhauben auf Hartholz Betten. Sie sind ad libitum pelletiert Nagerfutter und Leitungswasser zur Verfügung gestellt. </li><li> Mäuse sind erlaubt 7 Tage vor dem Beginn der Studie zu akklimatisieren. </li></ol> 2. Cells <ol><li> Die murine Urothelkarzinom-Zelllinie MB49 wird verwendet, um die Blase Tumoren zu erzeugen. </li><li> Die Zellen werden in Dulbecco modifiziertem Eagle-Medium mit 10% fötalem Rinderserum, 100 Einheiten / ml Penicillin und 100 Einheiten / ml Streptomycin kultiviert. </li><li> Die Kulturen werden bei 37 ° C und 5% CO 2 mit Medien ändert sich alle zwei vor drei Tagen. </li></ol> 3. Tierische orthotopen Tumormodell Die Instillation der Blase Tumor-Modell beschrieben ist hier eine Modifikation des Verfahrens von Gunther et al. 2 <ol><li> Anästhesie ist mit 2% Isofluran (MWI / VetOne, Meridian, ID, Kat. Nr. 501017) in Sauerstoff induziert und bei 0,75-1,25%. (Mehrere Tiere gleichzeitig behandelt werden können, was eine Erhöhung der Isofluran Ebenen erfordern.) Um Hornhaut Trocknung zu verhindern, werden die Augen mit einer künstlichen Tränen Augensalbe geschmiert. Körperwärme wird mit einem Heizkissen unter einem Papier drapieren gepflegt. </li><li> Das Fell des unteren Rückens ist abgeschnitten, um Kontakt mit dem Metall-Erdungsscheibe zu fördern und die rasierte Fläche ist mit electroconduction Gel beschichtet. </li><li> Jede Maus wird dann individuell in dorsalen recumbancy auf eine Metallplatte Erdungsplatte platziert, und die Blase katheterisiert transurethral mit einem geschmierten 24-Gauge-, 3 / 4 Zoll, over-the-Nadel IV-Katheter mit Mandrin entfernt. </li><li> Die Harnblase ist der Urin abgelassen und einem Platin-Draht wird durch den Katheter in die Blase zu verlassen freiliegende Drähte an jedem Ende des Katheters eingesetzt - etwa 2 mm auf der luminalen Ende des Katheters. Der Draht ist aus Platin, weil der Widerstand des Materials gegen Oxidation hergestellt. </li><li> Ein 2,5 W Strom wird dann in die Blase für ca. 0,5 Sekunden angewendet, indem Sie den äußeren Teil des Drahtes mit einem Elektrokauter Gerät auf die Blutgerinnung. </li><li> Der Draht wird dann an einen zweiten Standort in der Blase und weitere 2,5 W Bezahlung bewegt. </li><li> Der Draht wird entfernt, und 100 ul MB49-Zellen bei einer Konzentration von 1x10 6 Zellen / ml durch den Katheter in die Blase über einen 1ml Spritze zugetropft. </li><li> Die Spritzen sind links an der Katheter-Efflux der Zellsuspensionen zu verhindern, und der Katheter-Mäuse werden unter Narkose für 90 Minuten gehalten, damit die Zellen, um den Brennvorgang Seiten anhängen. </li><li> 100-200 ul sterilem Kochsalzlösung wird von IP Injektion jeder Maus vor der Verwertung verwaltet. </li><li> Am Ende der Inkubationszeit die Katheter entfernt und die Tiere dürfen auf dem Heizkissen erholen, bis aufrichtende Reflexe, zurückzukehren, an welcher Stelle die Mäuse ihre Käfige zurückgegeben werden. </li></ol> 4. Ultraschall <ol><li> Tumoren sind erlaubt, für 3-5 Tage wachsen, bevor die Mäuse Ultraschall-Untersuchung der Harnblase zu unterziehen, um das Tumorwachstum zu überwachen. </li><li> Für den ersten Ultraschall-Untersuchungen werden die Tiere betäubt, wie oben beschrieben. Mäuse sind in dorsalen recumbancy gelegt, und Bauchhaut wird durch Abschneiden der Pelz vorbereitet. </li><li> A 24 gauge IV Katheter transurethral in die Harnblase eingebracht. An diesem Punkt kann die Blase entleert und der Urin zur weiteren Analyse gesammelt. </li><li> 100 ul des sterilen Kochsalzlösung in die Blase durch den Katheter instilliert, um Blase Sichtbarkeit im Ultraschall zu verbessern. </li><li> Ultraschall Leitung Gel auf den Bauch aufgetragen und Sonographie durchgeführt, um die Blasenwand und Lumen zu beurteilen. </li></ol> 5. Repräsentative Ergebnisse: Tumorgewebe wird beginnen zu erscheinen 3-5 Tage nach der Inokulation. In unserer Erfahrung, in einer Gruppe von 12 Mäusen, haben 50% oder mehr routinemäßig eine nachweisbare Gewebemasse in der Blase (0,5 bis 1,0 ul, mit V = π / 6 · L · W · H) bei Tag fünf. Bei der Verwendung von MB 49 Zellen, entwickeln sich zu Tumoren weiter wachsen und Euthanasie wird von Tag 24 erforderlich sein, wenn die Mäuse unbehandelt bleiben. Sterbehilfe wird empfohlen, wenn Mäuse lethargisch oder kachektischen geworden, Anzeige zerzauste Fell, gebeugt und non-responsive Verhalten, oder wenn sie verlieren 20% ihres ursprünglichen Körpergewichts. &lt;/ P&gt; Die Tumoren wachsen schnell und werden nekrotische Zentren haben, nachdem sie eine Masse von 200 mg oder mehr, 3 angeblich, weil die Zellen in den Entwicklungsländern Tumor schneller als das, was sein kann durch den angiogenen Blutversorgung gewartet vermehren zu erreichen. Man kann auch indirekt überwachen Tumorprogression über das Niveau der hematurea, die vermutlich um eine schnelle Angiogenese in Tumoren zu entwickeln verwandt. Es wurde konsequent darauf hingewiesen, dass der Urin von unbehandelten tumortragenden Mäusen Blut am Tag 14 enthalten sind. Die Konzentration der roten Blutkörperchen erschien im Laufe der Zeit in diesen Mäusen zu erhöhen, und die Volumina der Urin gesammelt vermindert. Tumoren in mehr als 50 mg kann auch mit relativer Leichtigkeit ertastet werden, die typischerweise von 14 Tage nach der Inokulation.

<ol>
	<li>Institute of Laboratory Animal Resources. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Guide for the Care and Use of Laboratory Animals. , (1996).</li><li>Gunther, J. H., Jurczok, A., Wulf, T., Brandau, S., Deinert, I., Jocham, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimizing+syngeneic+orthotopic+murine+bladder+cancer+(MB49).">Optimizing syngeneic orthotopic murine bladder cancer (MB49).</a> Cancer Res. 59, 2834-2837 (1999).</li><li>Zhang, X., Atala, A., Godbey, W. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression-targeted+gene+therapy+for+the+treatment+of+transitional+cell+carcinoma.">Expression-targeted gene therapy for the treatment of transitional cell carcinoma.</a> Cancer Gene Ther. 15, 543-552 (2008).</li></ol>

Tierversuche wurden in Übereinstimmung mit den Richtlinien und Vorschriften her durch die Institutional Pflege und Nutzung des Ausschusses Tulane University gesetzt durchgeführt.

Diese minimal-invasive Methode zur Ermittlung orthotopen Blasentumoren bei weiblichen Mäusen kann auch für andere Zelltypen als MB49 angewendet werden, obwohl unterschiedliche Ergebnisse zu rechnen. Wenn weniger aggressive Tumorzellen verwendet werden, wird die Erhöhung der Leistung leicht (unter Verwendung von 0,5 W-Schritten auf die neue optimale etablieren) ergeben eine Fläche von mehr zugänglich Tumorzellen Anhang und mehr Blase Inkubationszeiten für die eingeflößt Zellen wird eine größere Menge an Ertrag ...

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<td>C57Bl/6J mice</td>
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an orthotopic model of murine bladder cancer

In diesem einfachen Verfahren werden Blasentumoren bei weiblichen Mäusen C57 durch den Einsatz von Katheterisierung, lokale Verätzungen und späteren Zelladhäsion gegründet. Nach ihrer Blasen transurethral sind katheterisiert und entleert werden die Tiere wieder katheterisiert, um das Einführen eines Platindrahtes in Blasen ohne Beschädigung der Harnröhre oder Blase zu ermöglichen. Die Katheter sind aus Teflon als Isolator für die Draht-, die elektrischen Strom in die Blase dirigiert eine Brandverletzung erstellen dienen. Ein Elektrokauter Einheit wird verwendet, um zu liefern 2,5 W bis das freie Ende des Drahtes, Abbrennen extrazellulären Schichten und bieten Anknüpfungspunkte für Karzinom-Zellen in Suspension in die Blase durch eine spätere Katheterisierung geliefert. Die Zellen bleiben in der Blase für 90 Minuten, nach denen der Katheter entfernt werden und die Blasen darf natürlich abtropfen lassen. Die Entwicklung des Tumors mittels Ultraschall überwacht. Besondere Aufmerksamkeit ist auf die Katheterisierung Technik im zugehörigen Video zu zahlen.

Watch this Scientific Journal Video about An Orthotopic Model of Murine Bladder Cancer at JoVE.com

An Orthotopic Model of Murine Bladder Cancer

Blasentumoren sind bei weiblichen Mäusen in eine minimal-invasive Weise durch Katheterisierung, lokale Verätzungen und späteren Haftung von Karzinomzellen, um den Brennvorgang Standorten etabliert.

Einem orthotopen Modell des Murine Blasenkrebs

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and ...

an-orthotopic-model-of-murine-bladder-cancer

Research

JoVE Journal

Medicine

15.8K Aufrufe.  Tulane University. Blasentumoren sind bei weiblichen Mäusen in eine minimal-invasive Weise durch Katheterisierung, lokale Verätzungen und späteren Haftung von Karzinomzellen, um den Brennvorgang Standorten etabliert.

Serie: An Orthotopic Model of Murine Bladder Cancer

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient management strategies. Orthotopic mouse models of oral cancer have been developed to facilitate the study of factors that impact invasion and serve as model system for evaluating anti-tumor therapeutics. In these systems, visualization of disseminated tumor cells within oral cavity tissues has typically been conducted by either conventional histology or with in vivo bioluminescent methods. A primary drawback of these techniques is the inherent inability to accurately visualize and quantify early tumor cell invasion arising from the primary site in three dimensions. Here we describe a protocol that combines an established model for squamous cell carcinoma of the tongue (SCOT) with two-photon imaging to allow multi-vectorial visualization of lingual tumor spread. The OSC-19 head and neck tumor cell line was stably engineered to express the F-actin binding peptide LifeAct fused to the mCherry fluorescent protein (LifeAct-mCherry). Fox1nu/nu mice injected with these cells reliably form tumors that allow the tongue to be visualized by ex-vivo application of two-photon microscopy. This technique allows for the orthotopic visualization of the tumor mass and locally invading cells in excised tongues without disruption of the regional tumor microenvironment. In addition, this system allows for the quantification of tumor cell invasion by calculating distances that invaded cells move from the primary tumor site. Overall this procedure provides an enhanced model system for analyzing factors that contribute to SCOT invasion and therapeutic treatments tailored to prevent local invasion and distant metastatic spread. This method also has the potential to be ultimately combined with other imaging modalities in an in vivo setting.

A comprehensive overview of the techniques involved in generating a mouse model of oral cancer and quantitative monitoring of tumor invasion within the tongue through multi-photon microscopy of labeled cells is presented. This system can serve as a useful platform for the molecular assessment and drug efficacy of anti-invasive compounds.

Multi-Photonen-Imaging der Tumorzellinvasion in einem orthotopen Mausmodell des oralen Plattenepithelkarzinoms

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

Forschung

Medizin

Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model

Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the understanding of the mechanics of tumor invasion, and direct intracerebral inoculation of tumor provides the opportunity of observing the invasive process in a physiologically appropriate environment2. As far as human brain tumors are concerned, the orthotopic models currently available are established either by stereotaxic injection of cell suspensions or implantation of a solid piece of tumor through a complicated craniotomy procedure3. In our technique we harvest cells from tissue culture to create a cell suspension used to implant directly into the brain. The duration of the surgery is approximately 30 minutes, and as the mouse needs to be in a constant surgical plane, an injectable anesthetic is used. The mouse is placed in a stereotaxic jig made by Stoetling (figure 1). After the surgical area is cleaned and prepared, an incision is made; and the bregma is located to determine the location of the craniotomy. The location of the craniotomy is 2 mm to the right and 1 mm rostral to the bregma. The depth is 3 mm from the surface of the skull, and cells are injected at a rate of 2 &mu;l every 2 minutes. The skin is sutured with 5-0 PDS, and the mouse is allowed to wake up on a heating pad. From our experience, depending on the cell line, treatment can take place from 7-10 days after surgery. Drug delivery is dependent on the drug composition. For radiation treatment the mice are anesthetized, and put into a custom made jig. Lead covers the mouse's body and exposes only the brain of the mouse. The study of tumorigenesis and the evaluation of new therapies for GBM require accurate and reproducible brain tumor animal models. Thus we use this orthotopic brain model to study the interaction of the microenvironment of the brain and the tumor, to test the effectiveness of different therapeutic agents with and without radiation.

The purpose of this article is to describe the use of an orthotopic glioblastoma model for chemoradiation studies. This article will go though cell processing, implanting, and radiotherapy of the mouse using an intracranial model.

Kombination Strahlentherapie in einem orthotopen Maus Brain Tumor-Modell

Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the ...

Development of Obliterative Bronchiolitis in a Murine Model of Orthotopic Lung Transplantation

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and standardized not only for the study of allo-rejection but also between syngeneic strains for examining mechanisms of ischemia-reperfusion injury after lung transplantation. Although the application of the rat and other large animal model 2 contributed significantly to the elucidation of these studies, the scope of those investigations is limited by the scarcity of knockout and transgenic rats. Due to no effective therapies for obliterative bronchiolitis, the leading cause of death in lung transplant patients, there has been an intensive search for pre-clinical models that replicate obliterative bronchiolitis. The tracheal allograft model is the most widely used and may reproduce some of the histopathologic features of obliterative bronchiolitis 3. However, the lack of an intact vasculature with no connection to the recipient's conducting airways, and incomplete pathologic features of obliterative bronchiolitis limit the utility of this model 4. Unlike transplantation of other solid organs, vascularized mouse lung transplants have only recently been reported by Okazaki and colleagues for the first time in 2007 5. Applying the basic principles of the rat lung transplant, our lab initiated the obliterative bronchiolitis model using minor histoincompatible antigen murine orthotopic single-left lung transplants which allows the further study of obliterative bronchiolitis immunopathogenesis6.

Obliterative bronchiolitis is the key impediment to the long-term survival of lung transplant recipients and the lack of a robust preclinical model precludes examining obliterative bronchiolitis immunopathogenesis. Unlike other solid organ transplants, vascularized mouse lung transplantation has only recently been developed. Here we show our independently developed obliterative bronchiolitis model after murine orthotopic single-lung transplantation.

Entwicklung einer Bronchiolitis obliterans in einem Mausmodell der orthotopen Lungentransplantation

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and ...

An Orthotopic Bladder Tumor Model and the Evaluation of Intravesical saRNA Treatment

We present a novel method for treating bladder cancer with intravesically delivered small activating RNA (saRNA) in an orthotopic xenograft mouse bladder tumor model. The mouse model is established by urethral catheterization under inhaled general anesthetic. Chemical burn is then introduced to the bladder mucosa using intravesical silver nitrate solution to disrupt the bladder glycosaminoglycan layer and allows cells to attach. Following several washes with sterile water, human bladder cancer KU-7-luc2-GFP cells are instilled through the catheter into the bladder to dwell for 2 hours. Subsequent growth of bladder tumors is confirmed and monitored by in vivo bladder ultrasound and bioluminescent imaging. The tumors are then treated intravesically with saRNA formulated in lipid nanoparticles (LNPs). Tumor growth is monitored with ultrasound and bioluminescence. All steps of this procedure are demonstrated in the accompanying video.

Establishing an orthotopic bladder tumor model to evaluate antitumor effects of intravesically delivered saRNA and monitoring tumor growth by ultrasound and bioluminescent imaging.

Einem orthotopen Blasentumor Modell und der Auswertung der intravesikalen Behandlung SÄRNA

We present a novel method for treating bladder cancer with intravesically delivered small activating RNA (saRNA) in an orthotopic xenograft mouse bladder ...

An Orthotopic Mouse Model of Anaplastic Thyroid Carcinoma

Several types of animal models of human thyroid carcinomas have been established, including subcutaneous xenograft and orthotopic implantation of cancer cells into immunodeficient mice. Subcutaneous xenograft models have been valuable for preclinical screening and evaluation of new therapeutic treatments. There are a number of advantages to using a subcutaneous model; 1) rapid, 2) reproducible, and 3) tumor establishment, growth, and response to therapeutic agents may be monitored by visual inspection. However, substantial evidence has shed light on the short-comings of subcutaneous xenograft models1-3. For instance, medicinal treatments demonstrating curative properties in subcutaneous xenograft models often have no notable impact on the human disease. The microenvironment of the site of xenographic transplantation or injection lies at the heart of this dissimilarity.
Orthotopic tumor xenograft models provide a more biologically relevant context in which to study the disease. The advantages of implanting diseased cells or tissue into their anatomical origin equivalent within a host animal includes a suitable site for tumor-host interactions, development of disease-related metastases and the ability to examine site-specific influence on investigational therapeutic remedies. Therefore, orthotopic xenograft models harbor far more clinical value because they closely reproduce human disease. For these reasons, a number of groups have taken advantage of an orthotopic thyroid cancer model as a research tool4-7.
Here, we describe an approach that establishes an orthotopic model for the study of anaplastic thyroid carcinoma (ATC), which is highly invasive, resists treatment, and is virtually fatal in all diagnosed patients. Cultured ATC cells are prepared as a dissociated cellular suspension in a solution containing a basement membrane matrix. A small volume is slowly injected into the right thyroid gland. Overall appearance and health of the mice are monitored to ensure minimal post-operative complications and to gauge pathological penetrance of the cancer. Mice are sacrificed at 4 weeks, and tissue is collected for histological analysis. Animals may be taken at later time-points to examine more advance progression of the disease. Production of this orthotopic mouse model establishes a platform that accomplishes two objectives: 1) further our understanding of ATC pathology, and 2) screen current and future therapeutic agents for efficacy in combating ATC.

Generation of an orthotopic mouse model of anaplastic thyroid carcinoma is described here. This technique employs surgical placement of human anaplastic thyroid cancer cells into the thyroid of immunodeficient mice, thus creating a more clinically relevant setting to study disease progression as well as screen innovative therapeutic interventions.

Ein orthotopen Mausmodell des Anaplastic Schilddrüsenkarzinom

Several types of animal models of human thyroid carcinomas have been established, including subcutaneous xenograft and orthotopic implantation of cancer ...

An Orthotopic Bladder Cancer Model for Gene Delivery Studies

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are needed. The tumor microenvironment can significantly influence tumor development and therapy response. It is therefore often desirable to grow tumor cells in the organ from which they originated. This protocol describes an orthotopic model of bladder cancer, in which MB49 murine bladder carcinoma cells are instilled into the bladder via catheterization. Successful tumor cell implantation in this model requires disruption of the protective glycosaminoglycan layer, which can be accomplished by physical or chemical means. In our protocol the bladder is treated with trypsin prior to cell instillation. Catheterization of the bladder can also be used to deliver therapeutics once the tumors are established. This protocol describes the delivery of an adenoviral construct that expresses a luciferase reporter gene. While our protocol has been optimized for short-term studies and focuses on gene delivery, the methodology of mouse bladder catheterization has broad applications.

Implantation of cancer cells into the organ of origin can serve as a useful preclinical model to evaluate novel therapies. MB49 bladder carcinoma cells can be grown within the bladder following intravesical instillation. This protocol demonstrates catheterization of the mouse bladder for the purpose of tumor implantation and adenoviral delivery.

Ein orthotopen Blasenkrebs-Modell für Gene Delivery Studies

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are ...

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules that regulate HSCs. In these transplant model systems, the function of HSCs is determined by the ability of these cells to engraft and reconstitute lethally irradiated recipient mice. Commonly, the donor cell contribution/engraftment is measured by antibodies to donor- specific cell surface proteins using flow cytometry. However, this method heavily depends on the specificity and the ability of the cell surface marker to differentiate donor-derived cells from recipient-originated cells, which may not be available for all mouse strains. Considering the various backgrounds of genetically modified mouse strains in the market, this cell surface/ flow cytometry-based method has significant limitations especially in mouse strains that lack well-defined surface markers to separate donor cells from congenic recipient cells. Here, we reported a PCR-based technique to determine donor cell engraftment/contribution in transplant recipient mice. We transplanted male donor bone marrow HSCs to lethally irradiated congenic female mice. Peripheral blood samples were collected at different time points post transplantation. Bone marrow samples were obtained at the end of the experiments. Genomic DNA was isolated and the Y chromosome specific gene, Zfy1, was amplified using quantitative Real time PCR. The engraftment of male donor-derived cells in the female recipient mice was calculated against standard curve with known percentage of male vs. female DNAs. Bcl2 was used as a reference gene to normalize the total DNA amount. Our data suggested that this approach reliably determines donor cell engraftment and provides a useful, yet simple method in measuring hematopoietic cell reconstitution in murine bone marrow transplantation models. Our method can be routinely performed in most laboratories because no costly equipment such as flow cytometry is required.

Determining donor cell engraftment presents a challenge in mouse bone marrow transplant models that lack well-defined phenotypical markers. We described a methodology to quantify male donor cell engraftment in female transplant recipient mice. This method can be used in all mouse strains for the study of HSC functions.

Mittels quantitativer real-time PCR zum Donor Cell Engraftment in einem Competitive Murine Bone Marrow Transplantation Modell fest

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%. New therapeutic options are critically needed and depend on improved understanding of pancreatic cancer biology. To better understand the interaction of cancer cells with the pancreatic microenvironment, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression. Luciferase-tagged pancreatic cancer cells are resuspended in Matrigel and delivered into the pancreatic tail during laparotomy. Matrigel solidifies at body temperature to prevent leakage of cancer cells during injection. Primary tumor growth and metastasis to distant organs are monitored following injection of the luciferase substrate luciferin, using in vivo imaging of bioluminescence emission from the cancer cells. In vivo imaging also may be used to track primary tumor recurrence after resection. This orthotopic model is suited to both syngeneic and xenograft models and may be used in pre-clinical trials to investigate the impact of novel anti-cancer therapeutics on the growth of the primary pancreatic tumor and metastasis.

Improved understanding of pancreatic cancer biology is critically needed to enable the development of better therapeutic options to treat pancreatic cancer. To address this need, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression using in vivo bioluminescence imaging.

Bioluminescent orthotopen Modell von Bauchspeicheldrüsenkrebs Progression

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%.  New therapeutic options are critically needed and depend on improved ...

An Orthotopic Murine Model of Human Prostate Cancer Metastasis

Our laboratory has developed a novel orthotopic implantation model of human prostate cancer (PCa). As PCa death is not due to the primary tumor, but rather the formation of distinct metastasis, the ability to effectively model this progression pre-clinically is of high value. In this model, cells are directly implanted into the ventral lobe of the prostate in Balb/c athymic mice, and allowed to progress for 4-6 weeks. At experiment termination, several distinct endpoints can be measured, such as size and molecular characterization of the primary tumor, the presence and quantification of circulating tumor cells in the blood and bone marrow, and formation of metastasis to the lung. In addition to a variety of endpoints, this model provides a picture of a cells ability to invade and escape the primary organ, enter and survive in the circulatory system, and implant and grow in a secondary site. This model has been used effectively to measure metastatic response to both changes in protein expression as well as to response to small molecule therapeutics, in a short turnaround time.

This orthotopic model of human prostate cancer allows for quantification of tumor size, circulating tumor cells, and formation of distinct metastasis to the lung. As cells must escape the primary organ, enter the blood stream, and implant into a secondary site, this model effectively recapitulates the scenario in humans.

Ein orthotopen Mausmodell der menschlichen Prostatakrebs-Metastasen

Our laboratory has developed a novel orthotopic implantation model of  human prostate cancer (PCa). As PCa death is not due to the primary tumor, but  ...

Minimally Invasive Establishment of Murine Orthotopic Bladder Xenografts

Orthotopic bladder cancer xenografts are the gold standard to study molecular cellular manipulations and new therapeutic agents&#160;in vivo. Suitable cell lines are inoculated either by intravesical instillation (model of nonmuscle invasive growth) or intramural injection into the bladder wall (model of invasive growth). Both procedures are complex and highly time-consuming. Additionally, the superficial model has its shortcomings due to the lack of cell lines that are tumorigenic following instillation. Intramural injection, on the other hand, is marred by the invasiveness of the procedure and the associated morbidity for the host mouse.
With these shortcomings in mind, we modified previous methods to develop a minimally invasive approach for creating orthotopic bladder cancer xenografts. Using ultrasound guidance we have successfully performed percutaneous inoculation of the bladder cancer cell lines UM-UC1, UM-UC3 and UM-UC13 into 50 athymic nude. We have been able to demonstrate that this approach is time efficient, precise and safe. With this technique, initially a space is created under the bladder mucosa with PBS, and tumor cells are then injected into this space in a second step. Tumor growth is monitored at regular intervals with bioluminescence imaging and ultrasound. The average tumor volumes increased steadily in in all but one of our 50 mice over the study period.
In our institution, this novel approach, which allows bladder cancer xenograft inoculation in a minimally-invasive, rapid and highly precise way, has replaced the traditional model.

The established technique to inoculate primary invasive orthotopic bladder cancer xenografts requires laparotomy and mobilization of the bladder. This procedure inflicts significant morbidity on the mice, is technically challenging and time-consuming. We therefore developed a high-precision, percutaneous approach utilizing ultrasound guidance.

Minimal Invasive Gründung von murinen orthotopen Blasen Xenografts

Orthotopic bladder cancer xenografts are the gold standard to study molecular cellular manipulations and new therapeutic agents&#160;in vivo. Suitable ...