Bu çalışma, Ulusal Bilim Vakfı KARİYER ödülü (CBET 0.846.395) ve Karşılaştırmalı Tıp Anabilim Dalı, Tulane Üniversitesi tarafından finanse edildi.

1. Hayvanlar <ol><li> Kadın C57BL/6J farelerde (Mus musculus) 17 fare virüsleri bulaşıcı ektoparazit, helmint endoparasites ve antikorlar ücretsiz nakliye önce satıcı tarafından doğrulanmaktadır. Fareler Tulane Üniversitesi Laboratuvar Hayvanları Bakım ve Kullanım Kılavuzu doğrultusunda AAALAC akredite hayvan tesisi içinde muhafaza edilir. 1 </li><li> Özellikle, fareler üzerinde bireysel parke yataklar havalandırılan polikarbonat cageswith microisolation filtre üstleri ev sahipliği grup (4-5 fare / kafes). Bu ad libitum pelet kemirgen gıda ve musluk suyu verilmektedir. </li><li> Fareler çalışma başlamadan 7 gün önce gelmesini izin verilir. </li></ol> 2. Hücreler <ol><li> Fare transizyonel hücreli karsinom hücre hattı MB49 mesane tümörleri üretmek için kullanılır. </li><li> Hücreler% 10 fetal sığır serumu, 100 ünite / ml penisilin ve 100 ünite / ml streptomisin ile desteklenmiş Dulbecco&#39;nun modifiye Eagle ortamında kültüre. </li><li> Kültürler 37 korunur ° C ve% 5 CO 2 medya ile 2-3 günde bir değişir. </li></ol> 3. Hayvan Ortotopik Tümör Modeli Mesane tümörü modeli burada açıklanan instilasyon Gunther ve arkadaşları tarafından açıklanan prosedürün bir değişiklik. 2 <ol><li> Anestezi oksijen izofluran% 2 (MWI / VetOne, Meridian, ID, Kedi # 501.017) ile indüklenen ve% 0,75-1,25 tutulur. (Birden fazla hayvanlar izofluran seviyelerinde bir artış gerektirecek, eş zamanlı olarak tedavi edilebilir.) Kornea buharlaşmayı önlemek için, göz bir suni gözyaşı oftalmik pomad ile yağlanır. Vücut ısı bir kağıt asmak altında ısıtılmış bir ped ile korunur. </li><li> Sırtın alt kürk metal topraklama plakası ile temas tanıtmak için kırpılır ve traş alan electroconduction jel ile kaplanmıştır. </li><li> Her fare sonra tek tek dorsal recumbancy metal bir topraklama plakası üzerine yerleştirilir ve mesane transüretral, 3 / 4 inç, over-the-iğne IV kateter kaldırıldı stile ile yağlanmış 24-ölçer kullanarak kateterize. </li><li> Mesane, idrar boşaltılır ve bir platin tel, kateterin her iki ucunda kablonun açıkta bırakarak mesane içine kateter sokulur - kateterin lumenal ucuna yaklaşık 2 mm. Oksidasyona malzeme direnci nedeniyle platin tel yapılır. </li><li> A 2.5 W akımı yaklaşık 0,5 saniye sonra pıhtılaşma için ayarlanmış bir elektrokoter ünitesi ile telin dış kısmına dokunarak mesane uygulanır. </li><li> Tel sonra ikinci bir konumu ve mesane uygulanan 2,5 W şarj taşınır. </li><li> Tel kaldırılır ve 1x10 6 hücre / ml, 1 ml şırınga yoluyla mesane içine kateter aracılığıyla aşılandığı bir konsantrasyon MB49 hücrelerinin 100 ul . </li><li> Şırıngalar hücre süspansiyonları efflux önlemek için kateterler eklenir ve kateterize farelerin hücreleri yanık siteler eklemek için izin vermek için 90 dakika için anestezi altında korunur bırakılır. </li><li> 100-200 ul steril serum fizyolojik kurtarma önce her fare IP enjeksiyon yoluyla uygulanır. </li><li> Kuluçka dönemi sonunda kateter çıkarılır ve hayvanlar dik refleksleri, farelerin evlerinde kafesleri iade edilir ve bu noktada, dönene kadar ısıtılmış pad üzerinde kurtarmak için izin verilir. </li></ol> 4. Ultrason <ol><li> Tümörler farelerde tümör büyümesini izlemek için mesane ultrasonla muayene önce 3-5 gün büyümeye izin verilir. </li><li> İlk ultrason muayenesi için, yukarıda açıklandığı gibi hayvanların anestezi. Fare dorsal recumbancy yerleştirilir ve karın deri kürk kırpma tarafından hazırlanır. </li><li> 24 gauge IV kateter mesane içine transüretral eklenir. Bu noktada, mesane boşaltılır ve daha fazla analiz için toplanan idrar olabilir. </li><li> 100 ul steril serum fizyolojik ultrasonda mesane görünürlüğünü artırmak için kateter yoluyla mesane içine aşılamış. </li><li> Ultrason iletim jel karın bölgesine uygulanan ve mesane duvar ve lümen değerlendirmek için ultrasonografi yapılır. </li></ol> 5. Temsilcisi Sonuçlar: Tümör dokusu aşılar 3-5 gün sonra ortaya çıkmaya başlar. Günde beş Deneyimlerimize göre, 12 farelerin bir grup,% 50 veya daha fazla rutin (1.0 ul, V = π / 6 · L · G · H kullanarak 0.5) mesane içinde algılanabilir bir doku kitlesi var. MB 49 hücreleri kullanılarak, tümör büyümeye devam edecek ve farelerin tedavi edilmezse giderseniz ötenazi günde 24 gerekecektir. Fareler uyuşuk veya kaşektik olunca Ötenazi önerilir hunching karıştırdı kürk, ve duyarlı olmayan davranış, ya da onların başlangıçtaki vücut ağırlığının% 20 kaybedersiniz. Görüntülemek./ P&gt; Bu tümörler hızlı büyür ve gelişmekte olan tümör hücrelerinin anjiyogenik kan temini ile hizmet ne olabilir daha hızlı çoğalırlar çünkü görünürde 200 mg veya daha fazla, 3 bir kitle ulaştıktan sonra nekrotik merkezleri olacak. Bir de dolaylı hematurea, muhtemelen gelişmekte olan tümörler içinde hızla anjiyogenez ile ilgili düzeyi ile tümör progresyonu izleyebilirsiniz. Bu sürekli tedavi edilmezse Tümör taşıyan farelere idrar 14 gün kan içerdiği kaydedildi. Kırmızı kan hücrelerinin konsantrasyonunun zamanla bu farelerde artırmak için ortaya çıkmış ve idrar hacimleri azalmış toplanan. 50 mg aşan tümörler de, tipik olarak 14 gün aşılar ile göreli kolaylıkla palpe edilebilir.

<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>

Tulane Üniversitesi Kurumsal Bakım ve Kullanım Kurulu tarafından belirlenen kurallara ve yönetmeliklere uygun olarak hayvanlar üzerinde deneyler yapıldı.

Bu minimal invaziv yöntem, dişi farelerde ortotopik mesane tümörleri kurulması, farklı sonuçlar beklenebilir rağmen daha MB49 diğer hücre türleri için de uygulanabilir. Daha az agresif tümör hücrelerinin kullanıldığı zaman, biraz watt (yeni optimum kurmak için 0,5 W artışlarını kullanarak) tümör hücre ek daha yumuşak bir alan verecektir ve aşılamış hücreleri için uzun mesane inkübasyon sürelerinde büyük miktarda verecektir yanık siteleri (yayınlanmamış veri) hücre eki. Bazı pr...

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<td>C57Bl/6J mice</td>
<td>&nbsp;</td>
<td>Jackson Laboratories</td>
<td>000664</td>
<td>3-5-wk-old</td>
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<td>Polycarbonate cages and ventilated rack</td>
<td>&nbsp;</td>
<td>Lab Products</td>
<td>Super Mouse 750</td>
<td>327mm x 190mm x 143mm</td>
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<td>Hardwood Bedding</td>
<td>&nbsp;</td>
<td>Harlan Teklad</td>
<td>Laboratory &#x2013;grade SaniChips</td>
<td>&nbsp;</td>
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<td>Rodent diet</td>
<td>&nbsp;</td>
<td>Harlan Teklad</td>
<td>Rodent Diet 2019</td>
<td>&nbsp;</td>
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<td>&nbsp;</td>
<td>MWI</td>
<td>009231</td>
<td>24 G x &frac34; in. </td>
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<td>Bovie Medical Corporation</td>
<td>Aaron 900</td>
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<td>&nbsp;</td>
<td>Sonosite</td>
<td>180 Plus (Vet)</td>
<td>&nbsp;</td>
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<td>MB49 cell line</td>
<td>&nbsp;</td>
<td>Anthony Atala, Wake Forest University Baptist Medical Center</td>
<td>N.A.</td>
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an orthotopic model of murine bladder cancer

Bu basit işlem, kateterizasyon, yerel koterizasyon, ve sonraki hücre adezyon yoluyla kadın C57 farelerde mesane tümörleri kurulur. Mesanesi transüretral kateterize ve süzülmüş sonra, hayvanlar, üretra veya mesane zarar vermeden tekrar mesanesi bir platin tel takılmasına izin kateterize. Kateterler, bir tel, bir yanık oluşturmak için mesane içine elektrik akımı yürütecek yalıtkan olarak hizmet Teflon yapılır. Ekstrasellüler katmanları uzakta yanan ve bir sonraki kateterizasyon ile mesane askıya teslim edilir karsinom hücreleri için eklenti sitelerini sağlayan bir elektrokoter ünitesi, tel maruz sonuna kadar 2.5W sunmak için kullanılır. Hücreler kateterler çıkarılır ve mesanesi doğal drenaj izin, 90 dakika sonra mesanede kalır. Tümör gelişimi ultrason ile takip edilir. Özel dikkat beraberindeki video kateterizasyon tekniği ödenir.

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

An Orthotopic Model of Murine Bladder Cancer

Kateterizasyon, yerel koterizasyon ve yanık sitelerine karsinom hücreleri sonraki yapışma ile minimal invaziv bir moda dişi farelerde mesane tümörleri kurulur.

Murin Mesane Kanseri bir Ortotopik Model

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 Görüntüleme.  Tulane University. Kateterizasyon, yerel koterizasyon ve yanık sitelerine karsinom hücreleri sonraki yapışma ile minimal invaziv bir moda dişi farelerde mesane tümörleri kurulur.

Video: 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.

Oral Skuamöz Hücre Karsinomu Ortotopik Fare Modeli Tümör Hücre Invasion Multi-foton Görüntüleme

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

Tıp

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.

Bir Ortotopik Fare Beyin Tümörü Modelinde Kombinasyon Radyoterapi

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.

Ortotopik Akciğer Transplantasyonu bir Mürin Modelinde Obliteratif Bronşiolit Gelişimi

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.

Bir Ortotopik Mesane Tümörü Model ve intravezikal Sarna Tedavisinin Değerlendirilmesi

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.

Anaplastik Tiroid Kanseri Ortotopik Fare Modeli

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.

Gene Teslim Çalışmaları Ortotopik Mesane Kanseri Modeli

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.

Rekabetçi bir mürin Kemik İliği Transplantasyon Modelinde Donör Hücre Aşı belirleme Kantitatif Gerçek-zamanlı PCR kullanarak

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.

Pankreas Kanseri İlerleme Bioluminescent Ortotopik Model

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.

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  ...

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.

Kemirgen Ortotopik Mesane ksenogratflarının Minimal İnvaziv kurulması

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