Name: a preclinical murine model of hepatic metastases
Uploaded: 2014-09-27T13:00:00.000+00:00
Duration: 6 min 51 s
Description: Watch this Scientific Journal Video about A Preclinical Murine Model of Hepatic Metastases at JoVE.com

KF ve KO KS ile birlikte ilk yazarları co edilir Bu çalışma AHPBA Kardeşliği (KS), NIH K23 CA148964-01 (LZ), Tıp Johns Hopkins Yüksekokulu Klinik Bilim İnsanı Ödülü (LZ), Virágh Vakfı ve Johns Hopkins Atla Virágh Pankreas Kanseri Merkezi (EMJ ve kısmen desteklenmiştir LZ), Ulusal Pankreas Vakfı (LZ), Lefkofsky Aile Vakfı (LZ), Gastrointestinal Kanserler P50 CA062924 yılında NCI SPORE (EMJ, LZ), Lustgarten Vakfı (LZ) ve Sol Goldman Pankreas Kanseri Merkezi hibeler (KS , BHE, LZ).

Tüm hayvan deneyleri Johns Hopkins Üniversitesi Hayvan Bakım ve Kullanım Kurulu kurallarına uyduğu, ve hayvanlar Laboratuarı Değerlendirme ve Bakımı Akreditasyon (AAALAC) Derneği kurallarına uygun olarak muhafaza edildi. Cerrahi işlem on beş saatte hava değişim minimum giren bir ameliyathanede steril koşullar altında gerçekleştirilir. Tüm cihazlar, işlem sırasında önce, her bir fare arasında% 70 etanol ile işlem ve tekrar otoklav ile sterilize edilir. Cerrahi prosedürü hayatta beklenmemektedir bütün fareler anestezi altında ise hala sonra servikal dislokasyon yapılmıştır 3 dakika boyunca CO2 altında kurban edilmiştir. 1. Hücre Hazırlanması <ol><li> 2 x10 7 hücre / ml fosfat tamponlu tuzlu su içinde kültürlenmiş hücrelerin tekrar Panc02 ve buz üzerinde tutun. </li></ol> 2. Fare Hazırlık <ol><li> Ketamin yönetmeXylazine (10 mg / kg) ile karıştırılmıştır E (100 mg / kg) intraperitoneal olarak 8-12 hafta bölünmüş dozlarda C57BL / 6 dişi fareleri kullanıldı. </li><li> Ayak tutam geri çekilme refleksi Fare tamamen uyuşturulduktan sağlamak için kaldırılmıştır olduğunu görmek için kontrol edin. </li><li> Tam fareler anestezi için gerektiği kadar ksilazin (10 mg / kg) ile karıştırılmıştır Ketamin ilave dozda (100 mg / kg) uygulayın. </li><li> Fareler anestezi altında iken kurumasını önlemek için, farelerin gözlerine Puralube Veteriner merhemi uygulanır. </li><li> Yaranın kirlenmesini önlemek için farelerin cerrahi alan tıraş. </li><li> Daha sonra iyot% 70 etanol ile kesi sol subcostal alanı hazırlayın. </li><li> Sterilliği korumak için, karın çevresinde bir cerrahi örtü yerleştirin. </li></ol> 3. Laparotomi <ol><li> Deri yoluyla ve bir neşter kullanılarak karın zarı aracılığıyla sol kulakta doğrultusunda sol subkostal kesi yapmak. </li><li> Eşzamanlı dijital p uygulayarak kesiden dalak ifadekesi kranial ve kaudal yönleri boyunca hava basıncı. </li><li> Dalak alt ucunda dalak, kan damarları bulun. </li><li> Dalak kutuplarda dalak, kan damarları zarar vermemek için dikkatli olmak dalak merkezinde iki Horizon orta boy ligatlayıcı klipleri koyarak dalak bölün. </li><li> Kirlenmeyi önlemek için geri periton içine dalak üst kutbunu yerleştirin. </li></ol> 4. Tümör Enjeksiyon <ol><li> Bir 26 G x 5/8 &quot;şırınga içine fosfat tamponlu tuzlu su, 150 ul hazırlayın. </li><li> Enjeksiyon sırasında her zaman dik bir pozisyonda şırınga muhafaza aynı enjektöre 100 Panc02 ul (2 x10 6) hücrelerini çizin. </li><li> % 70 etanol çözeltisi içinde iğne batırın. İğne sterilizasyonu sağlamak için% 70 etanol içinde daldırılır. </li><li> Şırınga her zaman dik tutulur emin olarak maruz hemispleen yavaşça hücreleri enjekte edilir. Enjektörler arasında şevge dalak altında hücrelerin enjekte edilmesini önlemek için dalak kapsülü ile her zaman dikkat edilmelidir. Dalak ve kan damarlarının bir beyazlatma enjeksiyonu üzerine dikkat edilmelidir. </li><li> Dalak yükseltmek ve dalak kan damarlarını tıkamak için dalak altında bir orta Horizon klibi geçerlidir. </li><li> Pankreas ve dalak gemilerin en distal yönüne küçük bir Horizon klibi uygulayın. </li><li> Horizon klipleri yukarıdaki keserek hemispleen gelen pankreas ve dalak gemilerin Arter. </li><li> Onun &#39;yan fare yerleştirin ve distile su ile kesi sulamak. </li></ol> Anestezi 5. Karın Kapanış ve Kurtarma <ol><li> 4-0 çalışan dikiş ile periton kapatın. </li><li> Cilt insizyon kapatmak için iki veya üç cilt klipleri uygulayın. </li><li> Cerrahi sonrası ağrıyı hafifletmek için 0.1 mg / kg buprenorfini veya 5-10 mg / kg Karprofen subkutan yönetmek. </li><li> Rec için bir ısıtma pedi üzerinde fare yerleştirinovery mobil kadar ve fare düzenli nefes desen göstermektedir. Hayvanlar sadece tam cerrahi kurtardıktan sonra diğer hayvanların şirkete iade edilir. </li></ol> 6. Nekropsi Sınav ve Karaciğer Hasat <ol><li> Cerrahi sonrası 30 ila 60 gün arasında, farenin metastaz klinik belirtileri göstermeye başlayacak ve ötenazi gerektirir. Insani ötenazi gerektiren metastatik hastalık belirtileri büyütülmüş karınlarını ve assit gelişimi bulunmaktadır. CO 2 Solunduğunda fareler Euthanize. </li><li> Teneffüs ardından, servikal çıkık gerçekleştirin. </li><li> Hayvan duyarlı olmayan olduğu tespit edildikten sonra, makas kullanılarak karın ortaya çıkarmak için karın zarı içinden bir kesik. </li><li> Forseps ve makas kullanarak, yavaşça karın dışına karaciğer kesti. </li><li> OCT bileşiği karaciğer yerleştirin ve flaş sıvı azot kullanılarak dondurmak. -80 ° C de karaciğer saklayın. Alternatif olarak, canlır gömülü 24 saat ve parafin oda sıcaklığında% 16 nötr tamponlu formalin içinde tespit edilebilir. </li></ol>

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Yazarlar ifşa ilgi hiçbir ilgili çatışmaları var.

Bir hemispleen enjeksiyon tekniği ile karaciğer, pankreas kanser metastazı bu klinik öncesi fare modeli, karaciğere metastatik hastalığı olan hastaların klinik ve immünolojik hastalıkların birçoğu aynaları tarif edilen birinci modelidir. Bu model, diğer sıçangil modellerinde fazla birkaç avantaj sunar. Farelerin sadece% 30 karaciğer ve metastaz oluşumu zamanlamasına metastazları geliştirir pankreas kanseri transjenik modelleri oldukça değişkendir farklı İlk olarak, bu model, farelerin% 100&#...

Pankreas duktal adenokarsinom (PDA) Amerika Birleşik Devletleri 1 kansere bağlı ölümlerin dördüncü önde gelen nedenidir. Metastatik PDA hastalar için beş yıllık sağkalım% 2-12 1&#39;dir. Pankreas kanseri için adjuvan kemoterapi ve bugün her zamankinden daha etkili olmasına rağmen, hala yeni tedaviler için umutsuz bir ihtiyaç duyulmaktadır. Yeni tedavilerin geliştirilmesi güvenilir klinik öncesi hayvan modellerinde gerektirir. , Deri altı tümör modelleri kullanmak için basit olmasına rağmen, dezavantajları, insan kanserlerinde görülen tümör ilerlemesi ve mikroçevrelerde metastaz taklit etmek için, bu tümörlerin yetersizlik içerir. Bu p53 ve Kras knock-onkojenik mutasyonların (KMG modeli) 2 ve bir soy YFP izleyicinin (PKCY model) KMG modeli içeren genetik olarak fare gibi genetik olarak modeller, 3 immunocompete doğal olarak oluşan tümörlerin çalışma için izin verirnt farenin. Buna ek olarak, tümör mikro değiştirilmemiş ve daha yakın insan tümör progresyonu görülene benzemektedir. Ne yazık ki, tümör gelişim zamanlaması zor deneysel tedavilerin çalışma yapar, bu model, içinde değişkendir. Ayrıca, bu farelerin geriye dönük melezleştirme zaman ve her zaman mümkün değildir, finansal bağlılık, önemli bir miktarda enerji gerektirmektedir. Son olarak, implante ksenogreft PDA fare modelleri değişmiş fareleri, ya da yoktur tümör mikroçevrelerde immünokompromize gerektirecek. Bunun bir sonucu olarak, bu klinik öncesi modellerinde ortaya deneysel tedavilerin etkinliği sıklıkla insan klinik deneylerinde 4 tekrarlanabilir değildir. Bu hemisplenectomy fare modeli C57BL / 6 farelerinin 5, 6 türetilmiş bir çok tümörijenik, metilkolantren kaynaklı pankreatik tümör hücre soyu olan Panc02 tümör hücrelerini kullanır. Ayrıca, diğer sıçangil PDA hücre çizgileri KMG mikrofon türetilmişE Bu modelde kullanılabilir. Schulick ve ark. Önce bir hemisplenectomy tekniğini geliştirmiş ve kolon kanseri metastazı 7 genden fare modeli oluşturduk. Bu hemisplenectomy modeli (Şekil 1) yeni terapötik ajanların 7-10 soruşturma için borç verme bağışıklığı farelerde tutarlı ve eşit tümör aşılama sunuyor.

<table><tbody><tr><td>Cell culture media and components</td><td></td><td></td><td>will vary depending on cell line</td></tr><tr><td>Phosphate Buffered Saline</td><td>Gibco by Life Technologies</td><td>10010-023</td><td></td></tr><tr><td>15 ml centrifuge tubes</td><td>Corning</td><td>430052</td><td></td></tr><tr><td>Curved Operating Scissor</td><td>Roboz Surgical Store</td><td>RS-6835</td><td></td></tr><tr><td>Micro Dissecting Graefe Forceps</td><td>Roboz Surgical Store</td><td>RS-5130</td><td></td></tr><tr><td>Needle holder (regular)</td><td>World Precision Instruments, Inc</td><td></td><td></td></tr><tr><td>3-0 or 4-0 suture</td><td></td><td></td><td>courtesy of dept of surgery, Johns Hopkins</td></tr><tr><td>Weck Horizon Open Ligating Clip Applier, small, angled</td><td>Teleflex</td><td>137085</td><td></td></tr><tr><td>Weck Horizon Open Ligating Clip Applier, medium, angled</td><td>Teleflex</td><td>237115</td><td></td></tr><tr><td>Weck Horizon medium ligating clips</td><td>Teleflex</td><td>002200</td><td></td></tr><tr><td>Weck Horizon small ligating clips</td><td>Teleflex</td><td>001200</td><td></td></tr><tr><td>Syringe, 1 cc, tb syringe, 3/8&quot; slip tip</td><td>Becton Dickinson</td><td>309625</td><td></td></tr><tr><td>Syringe, 1 cc, tb syringe, 5/8&quot; slip tip</td><td>Becton Dickinson</td><td>309597</td><td></td></tr><tr><td>9 mm Autoclip Applier</td><td>Mikron</td><td>427630</td><td></td></tr><tr><td>9 mm Autoclip</td><td>Becton Dickinson</td><td>427631</td><td></td></tr><tr><td>Heating pad</td><td>Sunbeam</td><td>CAT93</td><td></td></tr><tr><td>70% Ethanol</td><td></td><td></td><td>sold by numerous vendors</td></tr><tr><td>Ketamine Hydrochloride&amp;nbsp;</td><td>Hospira</td><td>22395DD</td><td></td></tr><tr><td>Xylazine hydrochloride</td><td>Sigma</td><td>X1251</td><td></td></tr></tbody></table>

a preclinical murine model of hepatic metastases

Sayısız fare modelleri, insan kanserleri çalışma ve kanser tedavisi ve kalkınma anlayışı ilerletmek için geliştirilmiştir. Burada, sinjenik mürin pankreas tümörü hücreleri hemispleen enjeksiyon ile karaciğer metastazlarının klinik öncesi, kemirgen pankreatik tümör modeli tarif edilmektedir. Bu model, karaciğere metastatik hastalığı olan hastalarda klinik durumların bir çoğu taklit eder. Fareler sürekli metastatik sürecinin incelenmesi, deneysel tedavi test ve tümör immünoloji araştırma için izin karaciğer metastazı gelişir.

Hemispleen içine enjekte edilen tümör hücreleri Panc02 tekniği doğru yapılması halinde akciğer ve karın zarı metastaz gözlenmez iken (Şekil 1) farelerin% 100&#39;ünde bir şekilde karaciğer metastazı. Bu teknik çok sayıda bağımsız deneylerde Panc02 hücreleri kullanılarak yüz üzerinde fareler üzerinde gerçekleştirilmiştir ve yeniden üretilebilir, tüm tedavi edilmemiş fareler hemisplenectomy prosedür (Şekil 2) takip eden 30 ila 60 gün, karaciğer metasta...

Watch this Scientific Journal Video about A Preclinical Murine Model of Hepatic Metastases at JoVE.com

A Preclinical Murine Model of Hepatic Metastases

Karaciğer metastazlarının klinik öncesi bir sıçangil modeli, hemispleen enjeksiyon tekniği ile gerçekleştirilir.

Karaciğer metastazları Klinik öncesi fare modeli

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

a-preclinical-murine-model-of-hepatic-metastases

Research

JoVE Journal

Medicine

28.6K Görüntüleme.  The Johns Hopkins University School of Medicine.  Karaciğer metastazlarının klinik öncesi bir sıçangil modeli, hemispleen enjeksiyon tekniği ile gerçekleştirilir. 

Video: A Preclinical Murine Model of Hepatic Metastases

Murine Bioluminescent Hepatic Tumour Model

This video describes the establishment of liver metastases in a mouse model that can be subsequently analysed by bioluminescent imaging. Tumour cells are administered specifically to the liver to induce a localised liver tumour, via mobilisation of the spleen and splitting into two, leaving intact the vascular pedicle for each half of the spleen. Lewis lung carcinoma cells that constitutively express the firefly luciferase gene (luc1) are inoculated into one hemi-spleen which is then resected 10 minutes later. The other hemi-spleen is left intact and returned to the abdomen. Liver tumour growth can be monitored by bioluminescence imaging using the IVIS whole body imaging system. Quantitative imaging of tumour growth using IVIS provides precise quantitation of viable tumour cells. Tumour cell death and necrosis due to drug treatment is indicated early by a reduction in the bioluminescent signal. This mouse model allows for investigating the mechanisms underlying metastatic tumour-cell survival and growth and can be used for the evaluation of therapeutics of liver metastasis. 

This article describes a procedure for the induction of orthotopic bioluminescent liver tumours in mice, and subsequent analysis of tumour growth confined to the liver using live whole body luminescence imaging.

Murin Bioluminescent Karaciğer Tümör Modeli

This video describes the establishment of liver metastases in a mouse model that can be subsequently analysed by bioluminescent imaging. Tumour cells are ...

Tıp

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.

İleri Hayvan Karaciğer Kolorektal Metastaz Model: Görüntüleme Teknikleri ve Metastatik Klonların Özellikleri

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

Kanser Araştırmaları

Surgical Procedures and Methodology for a Preclinical Murine Model of De Novo Mammary Cancer Metastasis

A rate-limiting aspect of transgenic mouse models of mammary adenocarcinoma is that primary tumor burden in mammary tissue typically defines study end-points. Thus, studies focused on elucidating mechanisms of late-stage de novo metastasis are compromised, as are studies examining efficacy of anti-cancer therapies targeting mediators of metastasis in the adjuvant setting. Numerous murine mammary cancer models have been developed via targeted expression of dominant oncoproteins to mammary epithelial cells yielding models variably mimicking histopathologic and transcriptome-defined breast cancer subtypes common in women1. While much has been learned regarding the biology of mammary carcinogenesis with these models, their utility in identifying molecules regulating growth of late-stage metastasis are compromised as mice are typically euthanized at earlier time points due to significant primary tumor burden. Moreover, since a significant percentage of women diagnosed with breast cancer receive adjuvant therapy after surgical resection of primary tumors and prior to presence of detectable metastatic disease, preclinical models of de novo metastasis are urgently needed as platforms to evaluate new therapies aimed at targeting metastatic foci. To address these deficiencies, we developed a murine model of de novo mammary cancer metastasis, wherein primary mammary tumors are surgically resected, and metastatic foci subsequently develop over a 115 day post-surgical period. This long latency provides a tractable model to identify functionally significant regulators of metastatic progression in mice lacking primary tumor, as well as a model to evaluate preclinical therapeutic efficacy of agents aimed at blocking functionally significant molecules aiding metastatic tumor survival and growth.

Surgical Procedures and Methodology for a Preclinical Murine Model of De Novo Mammary Cancer Metastasis

Pre-clinical models evaluating adjuvant therapy targeting breast cancer metastasis are lacking. To address this, we developed a murine model of de novo pulmonary mammary adenocarcinoma metastasis, wherein therapies administered in the adjuvant setting (post surgical resection of primary tumors) can be evaluated for efficacy in impacting previously seeded pulmonary metastases.

Klinik Öncesi Bir Murin Modeli İçin Cerrahi Prosedürler ve Metodoloji<emDe Novo</emMeme Kanseri Metastazı

A rate-limiting aspect of transgenic mouse models of mammary adenocarcinoma is that primary tumor burden in mammary tissue typically defines study ...

A Portal Vein Injection Model to Study Liver Metastasis of Breast Cancer

Breast cancer is the leading cause of cancer-related mortality in women worldwide. Liver metastasis is involved in upwards of 30% of cases with breast cancer metastasis, and results in poor outcomes with median survival rates of only 4.8 - 15 months. Current rodent models of breast cancer metastasis, including primary tumor cell xenograft and spontaneous tumor models, rarely metastasize to the liver. Intracardiac and intrasplenic injection models do result in liver metastases, however these models can be confounded by concomitant secondary-site metastasis, or by compromised immunity due to removal of the spleen to avoid tumor growth at the injection site. To address the need for improved liver metastasis models, a murine portal vein injection method that delivers tumor cells firstly and directly to the liver was developed. This model delivers tumor cells to the liver without complications of concurrent metastases in other organs or removal of the spleen. The optimized portal vein protocol employs small injection volumes of 5 - 10 &#956;l, &#8805; 32 gauge needles, and hemostatic gauze at the injection site to control for blood loss. The portal vein injection approach in Balb/c female mice using three syngeneic mammary tumor lines of varying metastatic potential was tested; high-metastatic 4T1 cells, moderate-metastatic D2A1 cells, and low-metastatic D2.OR cells. Concentrations of &#8804; 10,000 cells/injection results in a latency of ~ 20 - 40 days for development of liver metastases with the higher metastatic 4T1 and D2A1 lines, and &#62; 55 days for the less aggressive D2.OR line. This model represents an important tool to study breast cancer metastasis to the liver, and may be applicable to other cancers that frequently metastasize to the liver including colorectal and pancreatic adenocarcinomas.

A surgical procedure was developed to deliver mammary tumor cells to the murine liver via portal vein injection. This model permits investigation of late stages of liver metastasis in a fully immune competent host, including tumor cell extravasation, seeding, survival, and metastatic outgrowth in the liver.

Bir Portal Ven Enjeksiyon Modeli Karaciğer Metastazı Meme Kanseri Eğitim için

Breast cancer is the leading cause of cancer-related mortality in women worldwide. Liver metastasis is involved in upwards of 30% of cases with breast ...

An Oncogenic Hepatocyte-Induced Orthotopic Mouse Model of Hepatocellular Cancer Arising in the Setting of Hepatic Inflammation and Fibrosis

The absence of a clinically relevant animal model addressing the typical immune characteristics of hepatocellular cancer (HCC) has significantly impeded elucidation of the underlying mechanisms and development of innovative immunotherapeutic strategies. To develop an ideal animal model recapitulating human HCC, immunocompetent male C57BL/6J mice first receive a carbon tetrachloride (CCl4) injection to induce liver fibrosis, then receive histologically-normal oncogenic hepatocytes from young male SV40 T antigen (TAg)-transgenic mice (MTD2) by intra-splenic (ISPL) inoculation. Androgen generated in recipient male mice at puberty initiates TAg expression under control of a liver-specific promoter. As a result, the transferred hepatocytes become cancer cells and form tumor masses in the setting of liver fibrosis/cirrhosis. This novel model mimics human HCC initiation and progression in the context of liver fibrosis/cirrhosis and reflects the most typical features of human HCC including immune dysfunction.

Here, we describe the development of a clinically relevant murine model of liver cancer recapitulating the typical immune features of hepatocellular cancer (HCC).

Hepatik İnflamasyon ve Fibrozis Ortamında Ortaya Çıkan Hepatosellüler Kanserin Onkojenik Hepatositkaynaklı Ortotopik Fare Modeli

The absence of a clinically relevant animal model addressing the typical immune characteristics of hepatocellular cancer (HCC) has significantly impeded ...

Murine Model of Metastatic Liver Tumors in the Setting of Ischemia Reperfusion Injury

Liver ischemia and reperfusion (I/R) injury, a common clinical challenge, remains an inevitable pathophysiological process that has been shown to induce multiple tissue and organ damage. Despite recent advances and therapeutic approaches, the overall morbidity has remained unsatisfactory especially in patients with underlying parenchymal abnormalities. In the context of aggressive cancer growth and metastasis, surgical I/R is suspected to be the promoter regulating tumor recurrence. This article aims to describe a clinically relevant murine model of liver I/R and colorectal liver metastasis. In doing so, we aim to assist other investigators in establishing and perfecting this model for their routine research practice to better understand the effects of liver I/R on promoting liver metastases.

We describe in detail a clinically relevant colorectal cancer liver metastases (CRLM) tumor model and the influence of liver ischemia reperfusion (I/R) in tumor growth and metastasis. This model can help to better understand the mechanisms underlying surgery-induced promotion of liver metastatic growth.

İskemi Reperfüzyon Hasarı Ayarında Metastatik Karaciğer Tümörlerinin Murine Modeli

Liver ischemia and reperfusion (I/R) injury, a common clinical challenge, remains an inevitable pathophysiological process that has been shown to induce ...

Portal Vein Injection of Colorectal Cancer Organoids to Study the Liver Metastasis Stroma

Hepatic metastasis of colorectal cancer (CRC) is a leading cause of cancer-related death. Cancer-associated fibroblasts (CAFs), a major component of the tumor microenvironment, play a crucial role in metastatic CRC progression and predict poor patient prognosis. However, there is a lack of satisfactory mouse models to study the crosstalk between metastatic cancer cells and CAFs. Here, we present a method to investigate how liver metastasis progression is regulated by the metastatic niche and possibly could be restrained by stroma-directed therapy. Portal vein injection of CRC organoids generated a desmoplastic reaction, which faithfully recapitulated the fibroblast-rich histology of human CRC liver metastases. This model was tissue-specific with a higher tumor burden in the liver when compared to an intra-splenic injection model, simplifying mouse survival analyses. By injecting luciferase-expressing tumor organoids, tumor growth kinetics could be monitored by in vivo imaging. Moreover, this preclinical model provides a useful platform to assess the efficacy of therapeutics targeting the tumor mesenchyme. We describe methods to examine whether adeno-associated virus-mediated delivery of a tumor-inhibiting stromal gene to hepatocytes could remodel the tumor microenvironment and improve mouse survival. This approach enables the development and assessment of novel therapeutic strategies to inhibit hepatic metastasis of CRC.

Portal vein injection of colorectal cancer (CRC) organoids generates stroma-rich liver metastasis. This mouse model of CRC hepatic metastasis represents a useful tool to study tumor-stroma interactions and develop novel stroma-directed therapeutics such as adeno-associated virus-mediated gene therapies.

Karaciğer Metastazı Stromasını İncelemek için Kolorektal Kanser Organoidlerinin Portal Ven Enjeksiyonu

Hepatic metastasis of colorectal cancer (CRC) is a leading cause of cancer-related death. Cancer-associated fibroblasts (CAFs), a major component of the ...

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.

İnsan Prostat Kanser metastaz Ortotopik fare modeli

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  ...

Non-Invasive PET/MR Imaging in an Orthotopic Mouse Model of Hepatocellular Carcinoma

Preclinical experimental models of hepatocellular carcinoma (HCC) that recapitulate human disease represent an important tool to study tumorigenesis and evaluate novel therapeutic approaches. Non-invasive whole-body imaging using positron emission tomography (PET) provides critical insights into the in vivo characteristics of tissues at the molecular level in real-time. We present here a protocol for orthotopic HCC xenograft creation with and without hepatic artery ligation (HAL) to induce tumor hypoxia and the assessment of their tumor metabolism in vivo using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG) PET/magnetic resonance (MR) imaging. Tumor hypoxia could be readily visualized using the hypoxia marker [18F]FMISO, and it was found that the [18F]FMISO uptake was higher in HCC mice that underwent HAL than in the non-HAL group, whereas [18F]FDG could not distinguish tumor hypoxia between the two groups. HAL tumors also displayed a higher level of hypoxia-inducible factor (HIF)-1&#945; expression in response to hypoxia. Quantification of HAL tumors showed a 2.3-fold increase in [18F]FMISO uptake based on the standardized value uptake (SUV) approach.

Here, we present a protocol to create orthotopic hepatocellular carcinoma xenografts with and without hepatic artery ligation and perform non-invasive positron emission tomography (PET) imaging of tumor hypoxia using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG).

Hepatosellüler Karsinomun Ortotopik Fare Modelinde Non-İnvaziv PET/MR Görüntüleme

Preclinical experimental models of hepatocellular carcinoma (HCC) that recapitulate human disease represent an important tool to study tumorigenesis and ...

A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of cancer metastasis are still unclear. Animal models are essential for elucidating the mechanisms and for evaluating novel strategies for the treatment of metastatic cancers. Here, an in-depth description of a &#8220;patient-like&#8221; orthotopic syngeneic mouse model for exploring the mechanisms of metastasis of solid organ tumors is provided. The survival surgical implantation of BNL 1ME A.7R.1 mouse hepatocellular carcinoma cells directly into the liver (the organ of origin) of the inbred wild-type immune competent laboratory mouse strain, BALB/c is described. The success and reproducibility of this methodology recommends it for widespread use in elucidating the biological mechanisms of solid organ cancer metastasis.

The metastatic spread of cancer is the major cause of cancer-related deaths. We provide an in-depth description of our survival surgery methodology for establishing a &#8220;patient-like&#8221; orthotopic syngeneic mouse model system for studying the mechanisms of metastasis in solid organ tumors.

Bir &quot;Hasta gibi&quot; Ortotopik singenik Fare Modeli Hepatoselüler karsinomu metastazı

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of ...

Karaciğer metastazları Klinik öncesi fare modeli

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