이 연구는 NIH의 교내 프로그램에서 기금에 의해 지원됩니다.

I. 셀 준비 <ol><li> 세포 이식되는 매 5 생쥐 한 80% 합류 225cm 3 플라스크에 대해 10% FBS으로 DMEM의 합류로 성장하고 있습니다. </li><li> 세포의 미디어 벗어 대기음, 칼슘이나 마그네슘없이 PBS의 10 ML로 씻는다. 대기음 PBS 해제 세포. </li><li> 트립신 3 ML과 세포를 Trypsinize, 10-15 분 기다려야하고, 미디어의 15 ML에 트립신을 중화. </li><li> Pipet 50 ML 원추형으로 모든 세포와 미디어. 두 개의 커다란 flasks 한 50 ML 원추형으로 맞습니다. </li><li> 4 ° C.에서 7 분을위한 1,600 rpm으로 세포를 던가 완전히 뜨는 기음. 차가운 PBS의 10 ML에 세포를 Resuspend, 가볍게 아래로 pipetting 및하여 세포를 헹굴. 셀을 펠렛은 PBS로 균일하게 혼합되어 있는지 확인하십시오. 서스펜션은 구름이어야합니다. </li><li> 현재 셀 카운트는 보습 바로 앞에 달린 풀베는 날 카운터를 사용하여 수행됩니다. 세포는 4 ° C.에서 7 분을위한 1,600 rpm으로 다시 centrifuged있다 </li><li> 세포의 2 차 원심 분리 등 후PBS 1 ML의 해적 모든 PBS와 resuspend. </li><li> 세포는 4 ° C. 10 분 3,000 rpm으로 PBS의 1ml에 소용되는 PBS는 aspirated하고, PBS는 U87 세포를 5 μl 당 1x10 6으로 최종 농도를 조정하기 위해 추가됩니다, 다른 세포 라인은 최적의 종양 성장을위한 세포의 서로 다른 숫자와 집중을 필요로 할 수 있습니다. </li><li> 최종 볼륨과 집중을 얻은 후 전지 eppendorf 튜브의 얼음에 보관됩니다. </li></ol> II. 이종 이식 <ol class="ualpha"><li> 마우스를 준비 <ol><li> 마우스 케타민을 / Xylazine / 식염수의 주입 믹스를 사용하여 anesthetized 있습니다. 혼합물 120 케타민을, 16 밀리그램 / Rompun과 식염수와 Perl의 MG / kg이다. 생쥐는 개별적으로 무게와 자신의 체중을 기준으로 10g 당 0.1 ML 주어집니다. 마취는 1 ML의 주사기의 IP에 연결된 26G 바늘로 제공됩니다. 그것은 고통 응답을위한자는 단계에 도달하기 위해 생쥐에 대한 약 15 분 소요됩니다. 당신의 고통 응답을 확인하실 수 있습니다그들의 노상 강도를 곤란에 의한 생쥐. </li><li> 마우스가 완전히 anesthetized되면​​ 눈 연고가 말리는에서 그들을 방지하기 위해 그들의 눈에 관리합니다. </li><li> 마우스를 입 표시줄에서 노치의 앞에 윗니와 stereotaxic의 침대에 배치됩니다. 침대 후 stereotaxic에 배치됩니다. </li><li> 코 가드는 머리의 움직임을 방지하기 위해, 그리고 귀 바가 가볍게 생쥐의 귀 운하에 배치하고 마우스의 코 위에 배치됩니다. 두개골 골절 및 / 또는 호흡에서 마우스를 방지할 수 있으므로 코 표시줄이나 너무 꽉 귀 막대 중 하나 배치를하지 않도록주의하십시오. </li><li> 마우스의 머리가 stereotaxic에 진드기 부호를 사용하는 수준이 과정에서 도움이 있는지 확인합니다. </li><li> 준비하려면 마우스, 알코올 및 providine 요오드는 요오드의 알코올 및 3 애플 리케이션 3 응용 프로그램의 총 번갈아 마우스 머리에 후한 적용됩니다. 70%있는 알코올 패드 중 하나는 최종 응용 프로그램에 적용됩니다머리. </li></ol></li><li> 수술 <ol><li> 프로 시저에서 모든 악기 절차를 시작하기 전에 70 %의 에탄올에서 악기를 autoclaving 또는 소킹에 의해 어느 소독해야합니다. </li><li> 절개는 대각선으로 다시 두개골의 왼쪽 뒷부분에 오른쪽 눈 뒤에서 머리 위에 이루어집니다. 절개는 10mm ~ 15 mm 길이에 대해 여야합니다. </li><li> Q-팁을 사용하여 천천히 두개골을 노출하고 두개골과 피부 사이에 멤브레인 닦아 절개를 구분합니다. 과도한 출혈을 핏자국이 얼룩하고 시청 Q-팁을 사용하십시오. </li><li> bregma을 시각화하기 위해 두개골의 맨을 쉽게받을 수 있습니다. 해밀턴 주사기 포인트 사용 bregma 통해 위치를. 두개골을 뚫고되는 구멍의 정확한 위치는 2 밀리미터 권리와 bregma (그림 2)의 앞쪽에 1mm입니다. 멸균 구입 외과의 느낌 - 스쳐 마커는, 뚫고되는 구멍의 위치를​​ 표시하는 데 사용됩니다. 구멍이 가까이 위치하고있다는 점에 유의하십시오terior 봉합 라인. </li><li> 다음으로, 18G 바늘을 통해 핀 꽃병이나 수사관에 22G 바늘의 포인트를 사용하여 표시된 위치에서 두개골에 작은 구멍을 드릴 다운합니다. </li><li> 세포의 5 μl와 10 μl 유리 해밀턴 주사기를로드합니다. 해밀턴 주사기 그것에 부착된 고정 33G 바늘로 무딘 30G 갖춘 10 μl입니다. 그들을 pipetting 또는 부착된 25G 바늘로 1ml 주사기를 사용하여 중, 그들을로드하기 전에 세포를 섞어주십시오. 해밀턴 주사기가로드되면, 두개골에 구멍을 주선해 줄 수있는 팔을에 다시 넣습니다. </li><li> 바늘은 두개골과 수준되면 두개골의 표면에 바늘의 번트 엔드를 내려라, 그것이 두개골의 표면 아래 3 mm의 깊이에까지 바늘을 낮춥니다. </li><li> 바늘이 저하되면 그것은 두뇌에 영향을 미치는 바늘에서 어떤 큰 충격을 피하기 위해 2 분 동안 거기 남아있을 것이다. 세포를 이온 주입하면 6~8분의 총 분당 1 μl의 속도이며, 어떤 backflo주의w 세포의 마지막이 이식되고 나면, 모든 세포가 적응할 수 있도록 또 다른 2 분을 위치에 바늘을 떠나 </li><li> 두뇌에서 바늘을 제거하고, Q-팁을로 입구를 청소하십시오. 각 솔루션과 그 순서 배 50% Actone, 100 % EtOH, 그리고 PBS로 바늘과 주사기를 청소 진행합니다. </li><li> 두개골의 피부는이 시점에서 봉합받을 준비가되어 있습니다. 5-0를 사용하면 PBS는 약 3 바늘로 절개를 닫습니다. </li><li> 마우스가 열 패드에 깨어 있도록 허용하고, 필요한 경우 안과 연고를 다시 적용합니다. 자체의 움직임을 보여주는 이후 마우스는 새장에 반환됩니다 (즉 머리를 리프팅하거나 어깨를 이동). </li></ol></li></ol> III. 방사선 및 약물에 의한 IC 임플란트의 치료 <ol class="ualpha"><li> 방사선 치료 <ol><li> 세포주에 따라 생쥐의 치료 7 일 후 임플란트에 자리 5 소요될 수 있습니다. 우리 부서는 방사선 대접을 위해 orthovoltage 방사선 치료 장치를 사용하여, 표준. </li><li> 이러한 BLI (Bioluminescence 이미징), MRI 또는​​ 중부 표준시와 같은 이미징 전에 임의화 (그림 3A)에 종양을 시각화하는 데 사용할 수 있습니다. </li><li> 마우스는 이전과 케타민을 / Xylazine / 식염수 칵테일과 anesthetized이며, 눈 연고가 눈을에 적용됩니다. </li><li> 생쥐는 생쥐의 헤드 중심을 향해 가리 키도록하는 원형 바퀴처럼 구성된 맞춤 제작한 방사선 지그에 위치하고 있습니다. 생쥐의 신체는 리드와 방사선을 차단할 수 있습니다. </li><li> 약물 치료는 다른 배달 노선 즉, SQ, IP 및 IV를 통해 전이나 방사선 중 하나를 부여하실 수 있습니다. </li></ol></li></ol> IV. 결과 마우스가 마취 주사 후 45-60분 이내에 절차에서 깨어나한다. 마우스 difficultly 수술 후 깨어난했으며 시간이 1을 초과했습니다 ½ 시간을 수술 후 경우, 마우스는 복구할 수 없으며 안락사는 효과적인 대안이있을 수 있습니다. 만약 일오히려 과도한 출혈하거나 프로 시저 안락사 동안 시점에서 갈라진 두개골되어도 좋습니다. 모두 잘하고 세포주에 따라되면 절차가 이루어지는 후 종양 2-4 주 이내에 형성 시작합니다. intracranial 이종 이식 모델을 사용하면 치료 효능은 보통 생존 사후 주입 5로 측정됩니다. 그러나, 이것은 그대로 IC 모델 징후와 증상은 죽기 전에 발생합니다. 마우스가 발생할 수 있습니다 신체적 증상 중 일부는 hunched 자세, 체중 손실, 그리고 비활성입니다. 신경학적인 증상 중 일부는 압류 헤드 기울기와 균형의 손실있을 수 있습니다. 그림 3B는자가 사망률에서 종양의 얼룩 보여줍니다. 카플란 - 마이어의 줄거리는 데이터를 (그림 4) 평가하는 데 사용됩니다. 50 % 생존의 일 수를 계산 및 제어에 비해됩니다. 조합 그룹이 각각의 방사선 및 약물 무기의 추가보다 더 오래 살았다면 조합이 성공합니다. <img alt = &quot;그림 1&quot;SRC = &quot;/ files/ftp_upload/3397/3397fig1.jpg&quot;/&gt; 그림 1. Stoelting Stereotaxic 장비. <img alt="그림 2" src="/files/ftp_upload/3397/3397fig2.jpg" /> 그림 2. 마우스 두개골에 드릴 구멍의 위치의 도표. <img alt="그림 3A" src="/files/ftp_upload/3397/3397fig3A.jpg" /> GBM 주입 후 그림 3A. BLI 이미징. <img alt="그림 3B" src="/files/ftp_upload/3397/3397fig3B.jpg" /> 그림 3B. 사망률에서 H와 GBM 종양의 E. <img alt="그림 4" src="/files/ftp_upload/3397/3397fig4.jpg" /> 그림 4. PARP 억제제 7 일 게시물 수술과 주사 U87의 IC 이식의 결과.

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우리는 공개 할게 없다.

이러한 glioblastoma multiforme와 같은 악성 프로그램 gliomas은 가장 일반적인 기본 뇌종양을 대표하고 음침한 예후에게 4 있습니다. GBM에 의해 영향을 환자의 생존은 수술, 방사선 및 화학 요법 5의 진보에도 불구하고 (9-12 즉, 몇 달 후 진단) 지난 10 년 동안 거의 변함없이 남아있다. 세포가 인간의 gliomas의 histological 기능에 최대한 가깝게 유사합니다; 예측 가능한 성장이 있어야 glioma 치?...

<table border="1"><tbody><tr><td> 연구실 표준 Stereotaxic Instrutment </td><td> 마우스 및 쥐 어댑터를 Stoelting </td></tr><tr><td> Betadine </td><td> 백퍼센트 EtOH </td></tr><tr><td> 70% EtOH </td><td> 50 % 아세톤 </td></tr><tr><td> 6 &quot;Q-팁 </td><td> 알코올 면봉 </td></tr><tr><td> 해밀턴 주사기 80,008 </td><td> Ethicon 5-0 PD </td></tr><tr><td> 1701 SN 31-33G / 1 &quot;/ PT3 바늘 </td><td> 한 ML의 주사기 </td></tr><tr><td> 25G 바늘 </td><td> 생리대 </td></tr><tr><td> 인공 눈물 연고 </td><td> 아주 좋은 스쳐 영구 마커 </td></tr></tbody></table>

combination radiotherapy in an orthotopic mouse brain tumor model

Glioblastoma multiforme의 (GBM) 1 가장 일반적이고 공격적인 성인 뇌종양입니다. 최근에는 거기에 종양이 침략의 역학에 대한 이해에 상당한 진전되어, 그리고 종양의 직접 했습니다만 접종은 생리학적으로 적절한 환경 2의 침략 과정을 관찰의 기회를 제공하고있다. 마찬가지로 지금까지 인간의 뇌종양에 대해서라면 현재 사용 가능한 orthotopic 모델은 복잡한 craniotomy 절차 3을 통해 종양의 고체 부분의 세포 현탁액 또는 주입의 stereotaxic 주입에 의한 중 설립된다. 우리의 기술 우리는 두뇌에 직접 이식하는 데 사용되는 세포 현탁액을 만들 조직 배양에서 세포를 수확. 수술 시간은 약 30 분 정도이며, 마우스가 지속 외과 비행기에있을 필요로 주사용 마취제가 사용됩니다. 마우스 Stoetling (그림 1)에 의해 만들어진 stereotaxic 지그에 위치합니다. 고물에응급실은 외과 영역이 청소와 준비가되어 있습니다, 절개가 만들어졌다, 그리고 bregma는 craniotomy의 위치를​​ 결정하기 위해 위치하고 있습니다. craniotomy의 위치는 오른쪽으로 2mm와 bregma에 주동이의 1mm입니다. 깊이는 두개골의 표면에서 3mm이며, 세포, 2 μl 매 2 분 속도로 주입하고 있습니다. 피부는 5-0 PDS로 봉합하고, 마우스는 가열 패드에 깨워 허용됩니다. 우리의 경험에서 세포주에 따라 치료는 수술 후 7~10일에서 자리를 차지할 수 있습니다. 마약 배달은 약물 성분에 따라 좌우됩니다. 방사선 치료를 마우스 anesthetized 있으며, 지그 만든 사용자 정의에 투입. 리드는 마우스의 시체를 커버하고 마우스만을 두뇌를 제공합니다. tumorigenesis와 GBM에 대한 새로운 요법의 평가 연구는 정확하고 재현성 뇌종양 동물 모델을 필요로합니다. 따라서 우리는 effectiven을 테스트, 두뇌의 microenvironment과 종양의 상호 작용을 연구하기 위해이 orthotopic 뇌 모델을 사용방사선와의없이 다양한 치료 요원의 ESS.

Watch this Scientific Journal Video about Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model at JoVE.com

Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model

이 문서의 목적은 chemoradiation 연구를위한 orthotopic glioblastoma 모델의 사용을 설명하는 것입니다. 이 문서에서는 이온 주입, 세포 가공 불구하고 이동하고 intracranial 모델을 사용하여 마우스의 방사선 치료합니다.

Orthotopic 마우스 뇌종양 모델의 조합 방사선 치료

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

combination-radiotherapy-in-an-orthotopic-mouse-brain-tumor-model

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JoVE Journal

Medicine

16.3K 조회수.  National Cancer Institute. 이 문서의 목적은 chemoradiation 연구를위한 orthotopic glioblastoma 모델의 사용을 설명하는 것입니다. 이 문서에서는 이온 주입, 세포 가공 불구하고 이동하고 intracranial 모델을 사용하여 마우스의 방사선 치료합니다.

비디오: Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model

An Orthotopic Model of Murine Bladder Cancer

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and subsequent cell adhesion. After their bladders are transurethrally catheterized and drained, animals are again catheterized to permit insertion of a platinum wire into bladders without damaging the urethra or bladder. The catheters are made of Teflon to serve as an insulator for the wire, which will conduct electrical current into the bladder to create a burn injury. An electrocautery unit is used to deliver 2.5W to the exposed end of the wire, burning away extracellular layers and providing attachment sites for carcinoma cells that are delivered in suspension to the bladder through a subsequent catheterization. Cells remain in the bladder for 90 minutes, after which the catheters are removed and the bladders allowed to drain naturally. The development of tumor is monitored via ultrasound. Specific attention is paid to the catheterization technique in the accompanying video.

Bladder tumors are established in female mice in a minimally invasive fashion through catheterization, local cauterization, and subsequent adhesion of carcinoma cells to the burn sites.

Murine 방광 암의 Orthotopic 모델

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

연구

의학

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.

구강 편평 세포 암종의 Orthotopic 마우스 모델에서 종양 세포 침략의 다중 광자 이미징

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

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There is little knowledge about in situ, in vivo, tumor hypoxia during intracranial development of malignant brain tumors because of lack of efficient means to monitor it in these deep-seated orthotopic tumors. Bioluminescence imaging (BLI), based on the detection of light emitted by living cells expressing a luciferase gene, has been rapidly adopted for cancer research, in particular, to evaluate tumor growth or tumor size changes in response to treatment in preclinical animal studies. Moreover, by expressing a reporter gene under the control of a promoter sequence, the specific gene expression can be monitored non-invasively by BLI. Under hypoxic stress, signaling responses are mediated mainly via the hypoxia inducible factor-1&alpha; (HIF-1&alpha;) to drive transcription of various genes. Therefore, we have used a HIF-1&alpha; reporter construct, 5HRE-ODD-luc, stably transfected into human breast cancer MDA-MB231 cells (MDA-MB231/5HRE-ODD-luc). In vitro HIF-1&alpha; bioluminescence assay is performed by incubating the transfected cells in a hypoxic chamber (0.1% O2) for 24 hr before BLI, while the cells in normoxia (21% O2) serve as a control. Significantly higher photon flux observed for the cells under hypoxia suggests an increased HIF-1&alpha; binding to its promoter (HRE elements), as compared to those in normoxia. Cells are injected directly into the mouse brain to establish a breast cancer brain metastasis model. In vivo bioluminescence imaging of tumor hypoxia dynamics is initiated 2 wks after implantation and repeated once a week. BLI reveals increasing light signals from the brain as the tumor progresses, indicating increased intracranial tumor hypoxia. Histological and immunohistochemical studies are used to confirm the in vivo imaging results. Here, we will introduce approaches of in vitro HIF-1&alpha; bioluminescence assay, surgical establishment of a breast cancer brain metastasis in a nude mouse and application of in vivo bioluminescence imaging to monitor intracranial tumor hypoxia.

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model

Bioluminescence imaging of hypoxia inducible factor-1&alpha; activity is applied to monitor intracranial tumor hypoxia development in a breast cancer brain metastasis mouse model.

유방암 뇌 전이의 종양 Hypoxia 역학 생체내의 Bioluminescence 이미징에서는 마우스 모델에서

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There ...

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.

Orthotopic 방광 종양 모델 및 Intravesical saRNA 트리 트먼트의 평가

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.

역 형성 갑상선 암의 동소 마우스 모델

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.

유전자 전달 연구 동소 방광암 모델

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

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.

인간의 전립선 암 전이의 동소 쥐과 모델

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  ...

An Orthotopic Glioblastoma Mouse Model Maintaining Brain Parenchymal Physical Constraints and Suitable for Intravital Two-photon Microscopy

Glioblastoma multiforme (GBM) is the most aggressive form of brain tumors with no curative treatments available to date.
Murine models of this pathology rely on the injection of a suspension of glioma cells into the brain parenchyma following incision of the dura-mater. Whereas the cells have to be injected superficially to be accessible to intravital two-photon microscopy, superficial injections fail to recapitulate the physiopathological conditions. Indeed, escaping through the injection tract most tumor cells reach the extra-dural space where they expand abnormally fast in absence of mechanical constraints from the parenchyma.
Our improvements consist not only in focally implanting a glioma spheroid rather than injecting a suspension of glioma cells in the superficial layers of the cerebral cortex but also in clogging the injection site by a cross-linked dextran gel hemi-bead that is glued to the surrounding parenchyma and sealed to dura-mater with cyanoacrylate. Altogether these measures enforce the physiological expansion and infiltration of the tumor cells inside the brain parenchyma. Craniotomy was finally closed with a glass window cemented to the skull to allow chronic imaging over weeks in absence of scar tissue development.
Taking advantage of fluorescent transgenic animals grafted with fluorescent tumor cells we have shown that the dynamics of interactions occurring between glioma cells, neurons (e.g. Thy1-CFP mice) and vasculature (highlighted by an intravenous injection of a fluorescent dye) can be visualized by intravital two-photon microscopy during the progression of the disease.
The possibility to image a tumor at microscopic resolution in a minimally compromised cerebral environment represents an improvement of current GBM animal models which should benefit the field of neuro-oncology and drug testing.

We have established a cortical orthotopic glioblastoma model in mice for intravital two-photon microscopy that recapitulates the biophysical constraints normally at play during the growth of the tumor. A chronic glass window replacing the skull above the tumor enables the follow-up of the tumor progression over time by two-photon microscopy.

소성을 Glioblastoma의 마우스 모델은 뇌 실질 물리적 제약 조건을 유지하고의 intravital 두 광자 현미경에 적합

Glioblastoma multiforme (GBM) is the most aggressive form of brain tumors with no curative treatments available to date.
Murine models of this pathology ...

Tumor Treating Field Therapy in Combination with Bevacizumab for the Treatment of Recurrent Glioblastoma

A novel device that employs TTF therapy has recently been developed and is currently in use for the treatment of recurrent glioblastoma (rGBM). It was FDA approved in April 2011 for the treatment of patients 22 years or older with rGBM. The device delivers alternating electric fields and is programmed to ensure maximal tumor cell kill1.
Glioblastoma is the most common type of glioma and has an estimated incidence of approximately 10,000 new cases per year in the United States alone2. This tumor is particularly resistant to treatment and is uniformly fatal especially in the recurrent setting3-5. Prior to the approval of the TTF System, the only FDA approved treatment for rGBM was bevacizumab6. Bevacizumab is a humanized monoclonal antibody targeted against the vascular endothelial growth factor (VEGF) protein that drives tumor angiogenesis7. By blocking the VEGF pathway, bevacizumab can result in a significant radiographic response (pseudoresponse), improve progression free survival and reduce corticosteroid requirements in rGBM patients8,9. Bevacizumab however failed to prolong overall survival in a recent phase III trial26. A pivotal phase III trial (EF-11) demonstrated comparable overall survival between physicians&#8217; choice chemotherapy and TTF Therapy but better quality of life were observed in the TTF arm10.
There is currently an unmet need to develop novel approaches designed to prolong overall survival and/or improve quality of life in this unfortunate patient population. One appealing approach would be to combine the two currently approved treatment modalities namely bevacizumab and TTF Therapy. These two treatments are currently approved as monotherapy11,12, but their combination has never been evaluated in a clinical trial. We have developed an approach for combining those two treatment modalities and treated 2 rGBM patients. Here we describe a detailed methodology outlining this novel treatment protocol and present representative data from one of the treated patients.

A novel methodology that is employed for the treatment of recurrent glioblastomas is described. This treatment approach employs the application of alternating electric tumor treating fields (TTFields), known as TTF Therapy in combination with bevacizumab, a targeted agent that is currently FDA approved as monotherapy.

재발 성 아교 모세포종의 치료를 위해 베바 시주 맙과 조합하여 필드 요법 치료 종양

A novel device that employs TTF therapy has recently been developed and is currently in use for the treatment of recurrent glioblastoma (rGBM). It was FDA ...

Dynamic Contrast Enhanced Magnetic Resonance Imaging of an Orthotopic Pancreatic Cancer Mouse Model

Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) has been limitedly used for orthotopic pancreatic tumor xenografts due to severe respiratory motion artifact in the abdominal area. Orthotopic tumor models offer advantages over subcutaneous ones, because those can reflect the primary tumor microenvironment affecting blood supply, neovascularization, and tumor cell invasion. We have recently established a protocol of DCE-MRI of orthotopic pancreatic tumor xenografts in mouse models by securing tumors with an orthogonally bent plastic board to prevent motion transfer from the chest region during imaging. The pressure by this board was localized on the abdominal area, and has not resulted in respiratory difficulty of the animals. This article demonstrates the detailed procedure of orthotopic pancreatic tumor modeling using small animals and DCE-MRI of the tumor xenografts. Quantification method of pharmacokinetic parameters in DCE-MRI is also introduced. The procedure described in this article will assist investigators to apply DCE-MRI for orthotopic gastrointestinal cancer mouse models.

The goal of this protocol is to apply dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) for orthotopic pancreatic tumor xenografts in mice. DCE-MRI is a non-invasive method to analyze microvasculature in a target tissue, and useful to assess vascular response in a tumor following a novel therapy.