Gli autori ringraziano i membri del Dipartimento di Ostetricia e Ginecologia e Cancer Biology per i loro utili discussioni e assistenza tecnica. Questo studio è stato sostenuto da una società giapponese per la promozione della Scienza (JSP) Grant-in-Aid per la ricerca scientifica (15K15604, a H. Kajiyama).

Il protocollo di trattamento segue le linee guida per la sperimentazione animale adottato da Nagoya University. 1. Preparazione di sospensioni cellulari <ol><li> linea umana ovarico chiaro carcinoma delle cellule. <ol><li> Mantenere ES-2 celle 12 in terreno RPMI-1640 con siero 10% fetale bovino (FBS) e penicillina / streptomicina (penicillina: 100 unità / ml, streptomicina: 0,1 mg / ml). Cellule di coltura a 37 ° C in un incubatore umidificato CO 2 5%. </li><li> Raccogliere cellule in fase di crescita logaritmica in 0,05% tripsina / 0.02% soluzione di EDTA (tripsina / EDTA). <ol><li> In primo luogo, rimuovere i vecchi media dal recipiente di coltura cellulare, poi lavare le cellule una volta con PBS (PBS senza Ca 2 + / Mg 2+, 3 - 4 ml per 25 cm 2). </li><li> Successivamente, aggiungere 1 ml per 25 cm 2 di tripsina / EDTA. Ruotare il recipiente di coltura cellulare per coprire il monostrato cellulare con tripsina / EDTA. Dopo aver rimosso e scartando tripsina / EDTA, restituire il cellularecultura nave per l&#39;incubatore per 3-5 minuti o fino a quando le cellule sono staccato. </li><li> Esaminate le cellule al microscopio per assicurare le cellule vengono staccate dalla superficie del recipiente di coltura cellulare e galleggianti. </li><li> Aggiungere siero fresco contenente terreno di crescita per inattivare la tripsina (5 ml ogni 25 cm 2), e risospendere le cellule. Trasferire le cellule risospese in un tubo da 15 ml. </li><li> Centrifugare a 300 xg per 5 min. Eseguire centrifugazione sia a 4 ° C oa temperatura ambiente. Rimuovere con cautela il surnatante e cellule con terreno di coltura risospendere (5 ml ogni 25 cm 2). </li></ol></li><li> Contare le cellule (ad esempio, per esclusione trypan blu) poi risospendere in terreno di crescita ad una densità di 2 x 10 8 cellule / ml. Mettere le cellule sospese in ghiaccio. </li></ol></li><li> Scongelare congelati matrice extracellulare (ECM) a base di aliquote idrogel in un bagno di ghiaccio. </li><li> Aggiungere la sospensione cellulare per aliquote idrogel ECM basati con un rapporto di 1: 1 (finale cella concentration: 1 x 10 5 cellule / ml di idrogel di medio-ECM-based) e mescolare accuratamente. Posizionare le sospensioni cellulari preparate in un bagno di ghiaccio fino al momento dell&#39;uso. </li></ol> 2. ortotopico inoculazione con sospensioni cellulari NOTA: La chirurgia non richiede una suite dedicata. La chirurgia può essere eseguita su un banco di laboratorio in una zona della camera che è separata e ha attività minima. Una cappa laminare può anche essere usato. strumenti chirurgici sterili devono essere utilizzati e forbici non devono essere usati per fare incisioni cutanee. <ol><li> Utilizzare femminile topi nudi (BALB / c nu / nu) a 8 settimane di età per questo modello inoculazione ortotopico. </li><li> Innanzitutto, anestetizzare il mouse utilizzando inalazione con isoflurano (di 2 - 4%). Controllare la profondità anestetico verificando la mancanza di recesso su pizzico punta ferma. Dopo il mouse è anestetizzato, applicare una blanda pomata oftalmica sterile agli occhi per prevenire l&#39;essiccazione, se necessario. </li><li> Posizionare il mouse anestetizzato su un operating fase ventre rivolto verso il basso. Disinfettare l&#39;area chirurgica più volte sia con l&#39;alcol e uno scrub a base di clorexidina a base di iodio o. </li><li> Fare un ~ 2 cm incisione verticale vicino alla colonna vertebrale tra l&#39;arco costale destra e del femore. Utilizzare un telo chirurgico sterile intorno al sito di incisione. Fissare le superfici cutanee incise con morsetti per mantenere l&#39;apertura. Nota: Retrattori possono essere utilizzati al posto dei morsetti. </li><li> Successivamente, una seconda incisione (circa 1 cm) al peritoneo parietale sotto la prima incisione per aprire la cavità addominale. Bloccare le superfici addominali incise. Agganciare il tessuto grasso che circonda il tubo ovaio e Falloppio omolaterale con un morsetto per rimuovere l&#39;ovaio dalla cavità addominale. posizionare con cura il ovaio e tube di Falloppio in garza. Poi, posizionare il mouse ventre rivolto verso il basso sotto il microscopio da dissezione. </li><li> Posizionare la sospensione cellulare in una siringa da insulina (1/2 cc, 29 G) appena prima dell&#39;iniezione. iniettare con cautela la sospensione cellulare preparato (1- 2 ml) nel ovaio. Per molti sec, trattenere l&#39;ago all&#39;interno dell&#39;ovaio garantire gelificazione della idrogel ECM-based. Dopo gelificazione, le cellule tumorali iniettate rimangono all&#39;interno dell&#39;ovaio. </li><li> tornare con attenzione l&#39;ovaio nel corpo. Suturare la seconda incisione con la seta medici sterili, e quindi sigillare la pelle con clip ferita. </li><li> Dopo l&#39;intervento chirurgico, i topi saranno somministrati una analgesia (per esempio, meloxicam, intramuscolare (IM) o sottocutanea (SC), 0,2 mg / kg) per iniziale periodo di 24 ore. </li></ol> Il trattamento post-chirurgico 3. <ol><li> Mettere ogni topo in una gabbia separata su carta pulita con un impacco caldo per mantenere il caldo del mouse fino a quando non ha ripreso conoscenza sufficiente a mantenere decubito sternale. Poi, tornare gli animali ai loro gabbie per l&#39;allevamento e il monitoraggio. Monitorare i topi al giorno per 3 - 7 giorni dopo il trattamento chirurgico. </li><li> Il monitoraggio di topi post-operatoria <ol><li> In caso di segni di dolore, somministrare analgesiaSICS giornaliero (es, meloxicam, im o sc, 0,2 mg / kg). </li><li> Se i topi hanno segni di infezione (ad esempio, arrossamento, gonfiore, scarico), amministrare iniezione giornaliera con un reagente anti-infettivi (ad esempio, la penicillina, im, 100.000 UI / kg). </li></ol></li><li> Rimuovere i materiali di chiusura della pelle 10 - 14 giorni dopo il trattamento post-chirurgico. </li></ol> 4. Analisi dei tumori e delle metastasi NOTA: Il tumore si formerà alle iniettati ovaio 2 - 3 settimane dopo l&#39;inoculazione. <ol><li> Sacrificare i topi dal biossido di carbonio, e quindi raccogliere i campioni (ad esempio, tumori, metastasi, organi) 13-15 per ulteriori analisi 14,16-20. NOTA: Se un sistema di imaging in vivo è disponibile, cellule che producono enzimi che bioluminescenza (ad esempio, luciferasi) o la proteina fluorescente (ad esempio, GFP) possono essere utilizzati in questo protocollo. Questo protocollo modificato fornire informazioni utili perla dinamica della diffusione delle cellule tumorali dal tumore primario. </li></ol>

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I modelli animali sono essenziali per analizzare i meccanismi di sviluppo del tumore, la progressione, metastasi, e l&#39;efficacia dei farmaci contro le cellule tumorali. I topi recanti cellule tumorali ovariche umane essere già stati utilizzati come modello animale. Qui, descriviamo una procedura meno invasiva per la somministrazione di cellule tumorali ovariche in un sito ortotopico. Utilizzando questa procedura, inoculazione in ovaio attraverso un approccio retroperitoneale sulla fascia dorsale offre notevoli vanta...

Epiteliale carcinoma ovarico (EOC) rappresenta il più alto tasso di mortalità per cancro-correlata tra neoplasie ginecologiche 1. EOC è associato ad una prognosi sfavorevole, dovuto principalmente alla sua fine la sintomatologia, ed è spesso associata a più disseminations intraperitoneali e metastasi a distanza 2-4. diffusione peritoneale è un processo multi-step. In primo luogo, le cellule tumorali si staccano dalle lesioni primarie, e migrano nella cavità addominale. Quando le cellule tumorali attribuiscono al mesotelio peritoneale, cominciano a invadere i tessuti attraverso il mesotelio 5,6. Al fine di comprendere meglio la biologia del tumore (per esempio, la progressione del cancro e la risposta terapeutica), modelli murini offrono una ricchezza di informazioni. Un xenotrapianto con cellule tumorali umane è ampiamente usato per modelli murini in cui le cellule tumorali umane vengono inoculati per via sottocutanea, intraperitoneale, per via endovenosa, e ortotopicamente. Un modello animale con una ortotopicamente gtumore rafted può più efficiente e preciso generare risultati che riflettono l&#39;ambiente del tumore negli esseri umani rispetto ad un modello animale con un tumore heterotopically innestato. Pertanto, per molti tumori umani, modelli di trapianto ortotopico sono stati stabiliti 7-11. Qui, descriviamo l&#39;inoculazione ortotopico di cellule EOC umane attraverso un approccio retroperitoneale sulla fascia dorsale, che ha limitato l&#39;invasività rispetto alla inoculazione ventrale. Questa tecnica può fornire una varietà di informazioni utili su EOC, in particolare il meccanismo di diffusione intraperitoneale.

<table><tbody><tr><td>Matrigel matrix&amp;nbsp;</td><td>BD</td><td>354234</td><td>ECM-based hydrogel</td></tr><tr><td>Insulin syringe</td><td>TERUMO</td><td>SS-05M2913</td><td>1/2 cc, 29 G x 1/2&quot;</td></tr><tr><td>Suturing needle with suture</td><td></td><td></td><td>11 mm, 3/8, Nylon6-0</td></tr><tr><td>Reflex 7 mm Wound Clip Applier</td><td>CellPoint Scientific</td><td>204-1000</td><td></td></tr><tr><td>&amp;nbsp;Reflex 7 mm wound clips</td><td>CellPoint Scientific</td><td>203-1000</td><td></td></tr></tbody></table>

murine experimental model of original tumor development and peritoneal metastasis via orthotopic inoculation with ovarian carcinoma cells

carcinoma ovarico epiteliale (EOC) è associato ad una prognosi infausta perché mostra disseminazione peritoneale. Per migliorare la prognosi, è importante controllare la diffusione peritoneale. Tuttavia, non è ancora chiaro in che modo le cellule tumorali si staccano da lesioni primarie e attaccano al mesotelio. La creazione di un modello animale adeguato è necessario per acquisire una comprensione del meccanismo di diffusione peritoneale in vivo. In questo studio, si introduce il processo dalla iniezione locale di cellule EOC nella superficie ovarica murino allo sviluppo di metastasi, compreso il peritoneo e organi distanti. Nudo femminile topi (BALB / c nu / nu) a 8 settimane di età sono stati utilizzati. In un campo di vista microscopico, cellule EOC (1 x 10 5 cellule / ml di matrice medio extracellulare (ECM) a base di idrogel / ovaio unilaterale / topo) sono stati iniettati in ovaie murine attraverso un approccio retroperitoneale sulla fascia dorsale. Questo metodo proposto è un leprocedura invasiva ss per il mouse e riduce al minimo i danni per l&#39;ovaio. Qui, descriviamo i passi metodologici nello sviluppo della formazione del tumore originale e metastatico di EOC.

<html> Sei topi nudi avevano le loro ovaie inoculati con la chiara linea di cellule di carcinoma delle cellule umane ES-2, come descritto in questo protocollo. Come mostrato in figura 1A, dopo 2 settimane, 5 dei 6 topi aveva un tumore dell&#39;ovaio inoculato con ES-2 cellule, ma non c&#39;era tumore nell&#39;ovaio controlaterale senza inoculazione (usato come controllo). Inoltre, 2 dei 5 topi hanno mostrato più disseminazione peritoneale con ascite. Cellule metastatizza...

Watch this Scientific Journal Video about Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells at JoVE.com

Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells

Per chiarire il processo a più fasi di diffusione peritoneale del carcinoma ovarico, questo studio presenta un modello sperimentale murino di sviluppo del tumore originale e metastasi peritoneali tramite inoculazione ortotopico con queste cellule tumorali.

Modello murino sperimentale di sviluppo del tumore originale e peritoneale Metastasi<em&gt; via</em&gt; Ortotopico inoculazione con cellule dell&#39;ovaio Carcinoma

Epithelial ovarian carcinoma (EOC) is associated with a poor prognosis because it shows peritoneal dissemination. To improve the prognosis, it is ...

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Cancer Research

Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells

10.4K Views.  Bell Research Center for Reproductive Health and Cancer.  Per chiarire il processo a più fasi di diffusione peritoneale del carcinoma ovarico, questo studio presenta un modello sperimentale murino di sviluppo del tumore originale e metastasi peritoneali tramite inoculazione ortotopico con queste cellule tumorali. 

Video: Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells

An Orthotopic Model of Serous Ovarian Cancer in Immunocompetent Mice for in vivo Tumor Imaging and Monitoring of Tumor Immune Responses

Background: Ovarian cancer is generally diagnosed at an advanced stage where the case/fatality ratio is high and thus remains the most lethal of all gynecologic malignancies among US women 1,2,3. Serous tumors are the most widespread forms of ovarian cancer and 4,5 the Tg-MISIIR-TAg transgenic represents the only mouse model that spontaneously develops this type of tumors. Tg-MISIIR-TAg mice express SV40 transforming region under control of the Mullerian Inhibitory Substance type II Receptor (MISIIR) gene promoter 6. Additional transgenic lines have been identified that express the SV40 TAg transgene, but do not develop ovarian tumors. Non-tumor prone mice exhibit typical lifespan for C57Bl/6 mice and are fertile. These mice can be used as syngeneic allograft recipients for tumor cells isolated from Tg-MISIIR-TAg-DR26 mice.
Objective: Although tumor imaging is possible 7, early detection of deep tumors is challenging in small living animals. To enable preclinical studies in an immunologically intact animal model for serous ovarian cancer, we describe a syngeneic mouse model for this type of ovarian cancer that permits in vivo imaging, studies of the tumor microenvironment and tumor immune responses.
Methods: We first derived a TAg+ mouse cancer cell line (MOV1) from a spontaneous ovarian tumor harvested in a 26 week-old DR26 Tg-MISIIR-TAg female. Then, we stably transduced MOV1 cells with TurboFP635 Lentivirus mammalian vector that encodes Katushka, a far-red mutant of the red fluorescent protein from sea anemone Entacmaea quadricolor with excitation/emission maxima at 588/635 nm 8,9,10. We orthotopically implanted MOV1Kat in the ovary 11,12,13,14 of non-tumor prone Tg-MISIIR-TAg female mice. Tumor progression was followed by in vivo optical imaging and tumor microenvironment was analyzed by immunohistochemistry.
Results: Orthotopically implanted MOV1Kat cells developed serous ovarian tumors. MOV1Kat tumors could be visualized by in vivo imaging up to three weeks after implantation (fig. 1) and were infiltrated with leukocytes, as observed in human ovarian cancers 15 (fig. 2).
Conclusions: We describe an orthotopic model of ovarian cancer suitable for in vivo imaging of early tumors due to the high pH-stability and photostability of Katushka in deep tissues. We propose the use of this novel syngeneic model of serous ovarian cancer for in vivo imaging studies and monitoring of tumor immune responses and immunotherapies.

An Orthotopic Model of Serous Ovarian Cancer in Immunocompetent Mice for in vivo Tumor Imaging and Monitoring of Tumor Immune Responses

To study in vivo tumor growth and tumor microenvironment, we used a syngeneic and orthotopic mouse model of ovarian cancer in immunocompetent animals. We transduced a mouse tumor cell line (MOV1) with Katushka fluorescent protein (MOV1KAT) and here we show its orthotopic implantation in ovary and in vivo imaging.

Un modello ortotopico di carcinoma ovarico sieroso in topi immunocompetenti per In vivo Tumore Imaging e monitoraggio del tumore Risposte immunitarie

Background: Ovarian cancer is generally diagnosed at an advanced stage where the case/fatality ratio is high and thus remains the most lethal of all ...

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Immunologia e infezioni

An Orthotopic Mouse Model of Ovarian Cancer using Human Stroma to Promote Metastasis

Ovarian cancer is characterized by early, diffuse metastasis with 70% of women having metastatic disease at the time of diagnosis. While elegant transgenic mouse models of ovarian cancer exist, these mice are expensive and take a long time to develop tumors. Intraperitoneal injection xenograft models lack human stroma and do not accurately model ovarian cancer metastasis. Even patient derived xenografts (PDX) do not fully recapitulate the human stromal microenvironment as serial PDX passages demonstrate significant loss of human stroma. The ability to easily model human ovarian cancer within a physiologically relevant stromal microenvironment is an unmet need. Here, the protocol presents an orthotopic ovarian cancer mouse model using human ovarian cancer cells combined with patient-derived carcinoma-associated mesenchymal stem cells (CA-MSCs). CA-MSCs are stromal progenitor cells, which drive the formation of the stromal microenvironment and support ovarian cancer growth and metastasis. This model develops early and diffuses metastasis mimicking clinical presentation. In this model, luciferase expressing ovarian cancer cells are mixed in a 1:1 ratio with CA-MSCs and injected into the ovarian bursa of NSG mice. Tumor growth and metastasis are followed serially over time using bioluminescence imaging. The resulting tumors grow aggressively and form abdominal metastases by 14 days post injection. Mice experienced significant decreases in body weight as a marker of systemic illness and increased disease burden. By day 30 post injection, mice met endpoint criteria of &gt;10% body weight loss and necropsy confirmed intra-abdominal metastasis in 100% of mice and 60%-80% lung and parenchymal liver metastasis. Collectively, orthotopic engraftment of ovarian cancer cells and stroma cells generates tumors that closely mimic the early and diffuse metastatic behavior of human ovarian cancer. Furthermore, this model provides a tool to study the role of ovarian cancer cell: stroma cell interactions in metastatic progression.

Orthotopic engraftment of ovarian cancer cells mixed with human stromal cells provides a mouse model that exhibits rapid, diffuse metastatic behavior that is characteristic of human ovarian cancer. This model also allows for the study of tumor cell and stromal cell interactions, as well as their role in tumor progression and metastasis.

Ovarian cancer is characterized by early, diffuse metastasis with 70% of women having metastatic disease at the time of diagnosis. While elegant ...

Ricerca sul cancro

Intratibial Osteosarcoma Cell Injection to Generate Orthotopic Osteosarcoma and Lung Metastasis Mouse Models

Osteosarcoma is the most common primary bone cancer in children and adolescents, with lungs as the most common metastatic site. The five-year survival rate of osteosarcoma patients with pulmonary metastasis is less than 30%.Therefore, the utilization of mouse models mimicking the osteosarcoma development in humans is of great significance for understanding the fundamental mechanism of osteosarcoma carcinogenesis and pulmonary metastasis to develop novel therapeutics. Here, detailed procedures are reported to generate the primary osteosarcoma and pulmonary metastasis mouse models via&#160;intratibia injection of osteosarcoma cells. Combined with the bioluminescence or X-ray live imaging system, these living mouse models are utilized to monitor and quantify osteosarcoma growth and metastasis. To establish this model, a basement membrane matrix containing osteosarcoma cells was loaded in a micro-volume syringe and injected into one tibia of each athymic mouse after being anesthetized. The mice were sacrificed when the primary osteosarcoma reached the size limitation in the IACUC-approved protocol. The legs bearing osteosarcoma and the lungs with metastasis lesions were separated. These models are characterized by a short incubation period, rapid growth, severe lesions, and sensitivity in monitoring the development of primary and pulmonary metastatic lesions. Therefore, these are ideal models for exploring the functions and mechanisms of specific factors in osteosarcoma carcinogenesis and pulmonary metastasis, the tumor microenvironment, and evaluating the therapeutic efficacy in vivo.

The present protocol describes intratibia osteosarcoma cell injection to generate mouse models bearing orthotopic osteosarcoma and pulmonary metastasis lesions.

Iniezione di cellule di osteosarcoma intratibiale per generare modelli murini di osteosarcoma ortotopico e metastasi polmonari

Osteosarcoma is the most common primary bone cancer in children and adolescents, with lungs as the most common metastatic site. The five-year survival ...

Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of care consisting of surgical debulking and combination chemotherapy consisting of platinum and taxane compounds, almost 90% of patients recur within a few years. In these patients the development of chemoresistant disease limits the efficacy of currently available chemotherapy agents and therefore contributes to the high mortality. To discover novel therapy options that can target recurrent disease, appropriate animal models that closely mimic the clinical profile of patients with recurrent ovarian cancer are required. The challenge in monitoring intra-peritoneal (i.p.) disease limits the use of i.p. models and thus most xenografts are established subcutaneously. We have developed a sensitive optical imaging platform that allows the detection and anatomical location of i.p. tumor mass. The platform includes the use of optical reporters that extend from the visible light range to near infrared, which in combination with 2-dimensional X-ray co-registration can provide anatomical location of molecular signals. Detection is significantly improved by the use of a rotation system that drives the animal to multiple angular positions for 360 degree imaging, allowing the identification of tumors that are not visible in single orientation. This platform provides a unique model to non-invasively monitor tumor growth and evaluate the efficacy of new therapies for the prevention or treatment of recurrent ovarian cancer.

We describe a non-invasive animal imaging platform that allows the detection, quantification, and monitoring of ovarian cancer growth and recurrence. This intra-peritoneal xenograft model mimics the clinical profile of patients with ovarian cancer.

Modello murino per l&#39;imaging non invasivo per rilevare e monitorare Ovarian Cancer Ricorrenza

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of ...

Medicina

Stabilire un modello 3D ex vivo per l'interazione cancro ovarico-omento

An Ex Vivo Model of Ovarian Cancer Peritoneal Metastasis Using Human Omentum

Ovarian cancer is the deadliest gynecologic malignancy. The omentum plays a key role in providing a supportive microenvironment to metastatic ovarian cancer cells as well as immune modulatory signals that allow tumor tolerance. However, we have limited models that closely mimic the interaction between ovarian cancer cells and adipose-rich tissues. To further understand the cellular and molecular mechanisms by which the omentum provides a pro-tumoral microenvironment, we developed a unique 3D ex vivo model of cancer cell-omentum interaction. Using human omentum, we are able to grow ovarian cancer cells within this adipose-rich microenvironment and monitor the factors responsible for tumor growth and immune regulation. In addition to providing a platform for the study of this adipose-rich tumor microenvironment, the model provides an excellent platform for the development and evaluation of novel therapeutic approaches to target metastatic cancer cells in this niche. The proposed model is easy to generate, inexpensive, and applicable to translational investigations.

Autore in primo piano: Modello avanzato ex vivo per lo studio dell'interazione cancro-microambiente adiposo 

This protocol describes the establishment of a three-dimensional (3D) ex vivo model of cancer cell-omentum interaction. The model provides a platform for elucidating pro-tumor mechanisms within the adipose niche and for testing novel therapies.

Un modello ex vivo di metastasi peritoneali del carcinoma ovarico con omento umano

Ovarian cancer is the deadliest gynecologic malignancy. The omentum plays a key role in providing a supportive microenvironment to metastatic ovarian ...

In vivo Imaging and Therapeutic Treatments in an Orthotopic Mouse Model of Ovarian Cancer

Human cancer and response to therapy is better represented in orthotopic animal models. This paper describes the development of an orthotopic mouse model of ovarian cancer, treatment of cancer via oral delivery of drugs, and monitoring of tumor cell behavior in response to drug treatment in real time using in vivo imaging system. In this orthotopic model, ovarian tumor cells expressing luciferase are applied topically by injecting them directly into the mouse bursa where each ovary is enclosed. Upon injection of D-luciferin, a substrate of firefly luciferase, luciferase-expressing cells generate bioluminescence signals. This signal is detected by the in vivo imaging system and allows for a non-invasive means of monitoring tumor growth, distribution, and regression in individual animals. Drug administration via oral gavage allows for a maximum dosing volume of 10 mL/kg body weight to be delivered directly to the stomach and closely resembles delivery of drugs in clinical treatments. Therefore, techniques described here, development of an orthotopic mouse model of ovarian cancer, oral delivery of drugs, and in vivo imaging, are useful for better understanding of human ovarian cancer and treatment and will improve targeting this disease.

In vivo Imaging and Therapeutic Treatments in an Orthotopic Mouse Model of Ovarian Cancer

Orthotopic animal models of ovarian cancer replicate better human disease and therefore enhance our understanding of cancer progression and tumor response to therapy. A mouse model receives an intrabursal injection of luciferase-expressing ovarian tumor cells. Treatment is administered via oral gavage. Tumor growth is monitored by in vivo imaging system.

Imaging in vivo e trattamenti terapeutici in un modello di mouse ortotopico di cancro ovarico

Human cancer and response to therapy is better represented in orthotopic animal models.  This paper describes the development of an orthotopic mouse model ...

Biologia

Quantitation of Intra-peritoneal Ovarian Cancer Metastasis

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States. Mortality is due to diagnosis of 75% of women with late stage disease, when metastasis is already present. EOC is characterized by diffuse and widely disseminated intra-peritoneal metastasis. Cells shed from the primary tumor anchor in the mesothelium that lines the peritoneal cavity as well as in the omentum, resulting in multi-focal metastasis, often in the presence of peritoneal ascites. Efforts in our laboratory are directed at a more detailed understanding of factors that regulate EOC metastatic success. However, quantifying metastatic tumor burden represents a significant technical challenge due to the large number, small size and broad distribution of lesions throughout the peritoneum. Herein we describe a method for analysis of EOC metastasis using cells labeled with red fluorescent protein (RFP) coupled with in vivo multispectral imaging. Following intra-peritoneal injection of RFP-labelled tumor cells, mice are imaged weekly until time of sacrifice. At this time, the peritoneal cavity is surgically exposed and organs are imaged in situ. Dissected organs are then placed on a labeled transparent template and imaged ex vivo. Removal of tissue auto-fluorescence during image processing using multispectral unmixing enables accurate quantitation of relative tumor burden. This method has utility in a variety of applications including therapeutic studies to evaluate compounds that may inhibit metastasis and thereby improve overall survival.

Ovarian cancer metastasis is characterized by numerous diffuse intra-peritoneal lesions, such that accurate visual quantitation of tumor burden is challenging. Herein we describe a method for in situ and ex vivo quantitation of metastatic tumor burden using red fluorescent protein (RFP)-labeled tumor cells and optical imaging.

La quantificazione di intra-peritoneale del cancro ovarico metastasi

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States.  Mortality is due to diagnosis of 75% of ...

Intraperitoneal Injection of Cells: A Method of Delivering Cancer Cells into the Peritoneal Adipose Tissue of Murine Model

Source: Krishnan, V. et al. <a href="https://www.jove.com/t/52721">Approaches to Study Ovarian Cancer Metastatic Colonization of Milky Spot Structures in Peritoneal Adipose.</a> J. Vis. Exp. (2015)
In this video, we perform an intraperitoneal injection of ovarian cancer cell suspension in a mouse model to study the colonization of ovarian cancer cells in peritoneal adipose tissues and their subsequent metastasis.&#160;

Iniezione intraperitoneale di cellule: un metodo per fornire cellule tumorali nel tessuto adiposo peritoneale del modello murino

Source: Krishnan, V. et al. Approaches to Study Ovarian Cancer Metastatic Colonization of Milky Spot Structures in Peritoneal Adipose. J. Vis. Exp. 
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Modello murino sperimentale di sviluppo del tumore originale e peritoneale Metastasi

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Esplora Altri Video

Capitoli in questo video

Un modello ortotopico di carcinoma ovarico sieroso in topi immunocompetenti per In vivo Tumore Imaging e monitoraggio del tumore Risposte immunitarie

An Orthotopic Mouse Model of Ovarian Cancer using Human Stroma to Promote Metastasis

Iniezione di cellule di osteosarcoma intratibiale per generare modelli murini di osteosarcoma ortotopico e metastasi polmonari

Modello murino per l'imaging non invasivo per rilevare e monitorare Ovarian Cancer Ricorrenza

Un modello ex vivo di metastasi peritoneali del carcinoma ovarico con omento umano

Un modello ex vivo di metastasi peritoneali del carcinoma ovarico con omento umano

Un modello ex vivo di metastasi peritoneali del carcinoma ovarico con omento umano

Imaging in vivo e trattamenti terapeutici in un modello di mouse ortotopico di cancro ovarico

La quantificazione di intra-peritoneale del cancro ovarico metastasi

Iniezione intraperitoneale di cellule: un metodo per fornire cellule tumorali nel tessuto adiposo peritoneale del modello murino