Name: in vivo bioluminescent imaging of mammary tumors using ivis spectrum
Uploaded: 2009-04-29T00:00:00.000+00:00
Duration: 8 min 53 s
Description: Watch this Scientific Journal Video about In vivo Bioluminescent Imaging of Mammary Tumors Using IVIS Spectrum at JoVE.com

루시페라제 표현 암 세포 라인의 다양한는 마우스 모델에서 미리 임상 연구에 사용될 수 있습니다. 이러한 세포는 쉽게 선택 마커에 대한 필요가 표준 미디어 성장하는 병원체가없는 냉동 문화로 제공됩니다. 우리의 실험을 위해, 우리는 생체내의 유전자 발현이나 tumorgenesis의 광학 지표 역할을 루시페라제 유전자를 표현 4T1 - luc2 murine 내유 종양 세포 라인을 사용합니다. 우리는 비 invasively 기본 종양의 성장을 추적하는 루시페라제 표현을 사용하지만, 그들은 또한 전이성 병변을 찾아 모니터링하는 데 사용할 수 있습니다. 루시페라제 활동 주입하기 전에, 그리고이 작업을 수행하기 위해서는 검증해야 90% 합류 플라스크는 trypsinization에 의해 수확 후 계산됩니다. 수행하는 50,000 셀이 다음 microtiter 접시 및 시리얼 dilutions의 단일 우물에 적절하고 있습니다. Luciferin는 ML 당 150ug에있는 우물에 추가 2 분 동안 incubated입니다. microplate는 IVIS이나 표현 수준을 결정하는 발광 판 Reader에서 몇 군데 있습니다. 이 세포 라인은 초당 세포 당 6,500 광자까지 표현하고 있지만, 초당 셀 당 500 광자 이상 어떤 표현 수준은 생체내에서 성공적으로 몇 군데 있습니다. 이제 우리는 최적의 활동의 세포를 가지고, 우리는 피하 주입 단계로 진행할 수 있습니다. 종양의 최적의 검출을 용이하게하기 위해, 우리는 athymic immunocompromised 누드 마우스 변형을 사용하고 있습니다. 주사하기 전에, 동물 깊은 마취에 대한 효과 anesthetized 있습니다. 다음 우리가 100ul PBS에서 250,000 세포까지 주사할 수는 측면으로 subcutaneously 주입하고 있습니다. 1 ML의 주사기에있는 세포를로드하고 26 게이지 바늘을 첨부합니다. 포셉는 &quot;텐트&quot;하고 기지에 세포를 주입 함께 부드럽게 피부를 들어 봐요. 새로 주입된 세포는 즉시 몇 군데 있습니다. 각 이미지 세션에서 kg의 체중 당 Luciferin의 150mg의 총 후 복막 구멍 두개의 주사를 통해 관리합니다. 본 연구에서는 동물은 일관성있는 광자 유량을 보장하기 위해 Luciferin 주입 후 10 분 몇 군데 있습니다. 우리는 다음 단계에서 당신에게 보여 드리죠. Luciferin 동력학은 매일 다를 수 있습니다. 이 특정 모델에서 Luciferin 주입 후 10 분 측정하면 15 %의 다양성의 범위 내에서 결과를 주었다. 우리 실험에서는 우리는 다시 - thinned 요금 결합 소자 ° C 최대 감도를 달성하기 위해 -90으로 냉각을 사용하여 생체내 이미징 시스템에서 IVIS 스펙트럼을 사용합니다. 절대 quantitation를 지원하기 위해, 시스템 다운 시간 동안 어두운 요금을 측정하고 초기화하는 동안 자체 보정을 실행합니다. 이미징을 시작하려면, IVIS 시스템을 초기화 (한 번의 클릭) 및 실험을위한 이미징 매개 변수를 설정합니다. 몇 군데되는 동물의 숫자에 대한보기의 필드를 선택합니다. 최대 5 동물은 필수 마취 매니폴드를 사용하여 장비 유지하실 수 있습니다. 무대는 상수입니다 37 ° C 동물​​의 체온을 유지하기 위해. EKG 포트가 스트레스 절차 기간 동안 동물의 건강을 모니터링하기 위해 제공됩니다. 이미지 소프트웨어, 노출 시간, F - 중지하고 픽셀을 생활 비닝은 세포주의 표현 수준에 따라 최적화할 수 있습니다. 이러한 설정은 양적 결과에 영향을주지 않고 실험 도중 언제든지 변경할 수 있습니다. IVIS 흰 빛과 이미지를 중첩되는 정량 발광 생물이나 형광 신호, 아래에있는 동물의 사진 이미지를 얻습니다. 발광 생물 신호는 초당 광자로 표현하고 강도지도로 표시됩니다. 이미지 디스플레이는 quantitation에 영향을주지 않고 이미지에 최적의 콘트라스트와 해상도를 제공하기 위해 조정됩니다. 세포에서 발광이 관심을 도구의 영역을 사용하여 주사의 사이트에서 측정할 수 있습니다. 측정 데이터는 저장하거나 분석을 위해 내보낼 수있는 이미지 캡처에 관한 모든 실험 매개 변수와 함께 테이블에 표시됩니다 여러 이미지를 획득하여 실험의 특성에 따라 초 개월에 걸쳐 종단 연구에 비교할 수 있습니다. 우리는 시간 제로에서 종양에서 광자 자속을 측정하고 격주 간격으로 이미지와 함께 4 주 동안 모니터합니다. 종양에서 광자 유량은 bioluminescence는 종양의 크기와 직접 연결합니다 그래서 루시페라제을 표현하는 라이브 세포의 수에 비례합니다. 오일 포스트 주입에 종양이 아직 만져서 알 수있는 것이 아니라 세포가 bioluminescence과 종양이 적극적으로 성장을 보아입니다 통해 계량하실 수 있습니다. 이 단계에서는 bioluminescence 신호는 훨씬 stronge입니다R. 노출 시간, F - 중지하고 픽셀 비닝는 이미지가 명확하고 카메라가 포화되지 않도록 조정할 수 있습니다. 이러한 측정은 실험에 이전 이후 수집들에게 비교할 수 있도록 IVIS 자동으로 집광의 변화에​​ 대한 보상. 칠일에 게시 주입 종양 측정은 이미 데이터가 7 일 발생한 것은 이번이 처음과 bioluminescence에 대한 만져서 알 수있는 수 있습니다. 28일 포스트 주입에서 종양 괴사되고 있으며, 세포는 죽지 시작합니다. 캘리퍼스 측정하여 예상 종양 크기는 appreciably 변경되지 않습니다하지만, 종양의 발광은 세포 죽음을 나타내는 줄어 듭니다. 인도적 종점에 도달할 때까지 캘리퍼스 측정 및 bioluminescence 측정 계속하실 수 있습니다. hypoxia 또는 치료 정권에 의한 종양 괴사들은 종양 질량을 감소하지 않더라도 감소 bioluminescence로 표시됩니다.

나는 시각 실험이 문서의 저널을 생산하기 위해 수행 모든 실험이 기관 애니멀 케어 승인한 동물 프로토콜 060을 준수 수행 있다고 확신하고 칼리퍼 생명 과학에서위원회 (IACUC), 알라 메다 CA 재 김 박사를 사용하고 싶습니다 . D., IACUC 위원장, 칼리퍼 생명 과학.

in vivo bioluminescent imaging of mammary tumors using ivis spectrum

4T1 마우스 내유 종양 세포는 15~20일에 만져서 알 수있는 종양을 형성 뉴 / 뉴 생쥐의 하위 cutaneously 이식 수 있습니다. 이 이종 이식 종양 모델 시스템 putative antitumor 화합물의 생체내 평가 미리 임상을 위해 중요합니다. 4T1 세포 라인은 constitutively 반딧불 루시페라제 유전자 (luc2)을 표현하기 위해 설계되었습니다. 4T1 - luc2 종양을 나르는 생쥐는 종양 IVIS 스펙트럼 같은 민감한 광학 이미징 시스템을 사용하여 모니터링할 수있는 영상 조명 신호를 방출 Luciferin로 주입하는 경우. 종양에서 광자 플럭스은 발광 세포의 개수에 비례하여 신호가 종양 성장과 발전을 모니터 측정할 수 있습니다. IVIS는 발광 생물 신호 및 종단 연구의 절대 quantitation이 양적 결과를 손상시키지 않고 몇 개월 동안 신호 크기의 여러 명령을 통해 수행할 수 있도록 조정합니다. 종양 크기가 기존의 물리적인 방법으로 만져서 알 수있는 또는 측정되기 전에 종양 성장 bioluminescence으로 며칠 동안 모니터링할 수 있습니다. 이것은 급속한 모니터링 종양 발달 초기에 이벤트에 대한 통찰력을 제공하거나 짧은 실험 절차를 초래할 수 있습니다. 종양 세포 죽음과 hypoxia 또는 약물 치료에 의한 괴사는 발광 생물 신호의 감소에 의해 초기 표시됩니다. 이 세포 죽음을 물리적인 방법으로 측정으로 종양의 크기 감소에 의해 동반되지 않을 수도 있습니다. 종양 괴사의 초기 이벤트를 볼 수있는 능력은 치료 요원의 선택과 개발에 상당한 영향을 미치고 있습니다. IVIS를 사용하여 종양 성장의 양적 영상은 정확한 quantitation를 제공하고 결과를 생성하기 위해 실험 과정을 가속.

Watch this Scientific Journal Video about In vivo Bioluminescent Imaging of Mammary Tumors Using IVIS Spectrum at JoVE.com

In vivo Bioluminescent Imaging of Mammary Tumors Using IVIS Spectrum

루시페라제 표현 내유 종양 세포는 생쥐의 subcutaneously 이식 및 비 invasively 종단 연구에 종양이 성장과 발전을 모니터 광학 이미징을 사용하여 시각입니다.

IVIS 스펙트럼을 사용하여 내유 종양의 생체내 이미징에서 발광 생물

4T1 mouse mammary tumor cells can be implanted sub-cutaneously in nu/nu mice to form palpable tumors in 15 to 20 days. This xenograft tumor model system ...

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Research

JoVE Journal

Biology

In vivo Bioluminescent Imaging of Mammary Tumors Using IVIS Spectrum

47.0K 조회수.  Caliper Life Sciences.  中文 

비디오: In vivo Bioluminescent Imaging of Mammary Tumors Using IVIS Spectrum

Establishing Intracranial Brain Tumor Xenografts With Subsequent Analysis of Tumor Growth and Response to Therapy using Bioluminescence Imaging

Transplantation models using human brain tumor cells have served an essential function in neuro-oncology research for many years. In the past, the most commonly used procedure for human tumor xenograft establishment consisted of the collection of cells from culture flasks, followed by the subcutaneous injection of the collected cells in immunocompromised mice. Whereas this approach still sees frequent use in many laboratories, there has been a significant shift in emphasis over the past decade towards orthotopic xenograft establishment, which, in the instance of brain tumors, requires tumor cell injection into appropriate neuroanatomical structures. Because intracranial xenograft establishment eliminates the ability to monitor tumor growth through direct measurement, such as by use of calipers, the shift in emphasis towards orthotopic brain tumor xenograft models has necessitated the utilization of non-invasive imaging for assessing tumor burden in host animals. Of the currently available imaging methods, bioluminescence monitoring is generally considered to offer the best combination of sensitivity, expediency, and cost. Here, we will demonstrate procedures for orthotopic brain tumor establishment, and for monitoring tumor growth and response to treatment when testing experimental therapies.

Luciferase-modified human brain tumor xenografts can be established intracranially in athymic mice, with subsequent monitoring of tumor growth and response to therapy using bioluminescence imaging. In combination with survival analysis, bioluminescence monitoring is an essential research tool for pre-clinical testing of therapies being considered for treating brain tumors.

Bioluminescence 이미징을 사용하여 종양 성장과 치료에 반응 이후 분석 Intracranial 뇌종양의 Xenografts 구축

Transplantation models using human brain tumor cells have served an essential function in neuro-oncology research for many years. In the past, the most ...

연구

신경과학

Monitoring Tumor Metastases and Osteolytic Lesions with Bioluminescence and Micro CT Imaging

Following intracardiac delivery of MDA-MB-231-luc-D3H2LN cells to Nu/Nu mice, systemic metastases developed in the injected animals. Bioluminescence imaging using IVIS Spectrum was employed to monitor the distribution and development of the tumor cells following the delivery procedure including DLIT reconstruction to measure the tumor signal and its location.
Development of metastatic lesions to the bone tissues triggers osteolytic activity and lesions to tibia and femur were evaluated longitudinally using micro CT. Imaging was performed using a Quantum FX micro CT system with fast imaging and low X-ray dose. The low radiation dose allows multiple imaging sessions to be performed with a cumulative X-ray dosage far below LD50. A mouse imaging shuttle device was used to sequentially image the mice with both IVIS Spectrum and Quantum FX achieving accurate animal positioning in both the bioluminescence and CT images. The optical and CT data sets were co-registered in 3-dimentions using the Living Image 4.1 software. This multi-mode approach allows close monitoring of tumor growth and development simultaneously with osteolytic activity.

An experimental mouse model of bone metastasis was established following intracardiac delivery of luciferase expressing mammary tumor cells. Tumor development and resulted osteolytic lesion were monitored longitudinally with bioluminescence and micro CT imaging.

Bioluminescence 및 마이크로 중부 표준시 이미징과 함께 종양의 Metastases 및 Osteolytic 조직 손상을 모니터링

Following intracardiac delivery of MDA-MB-231-luc-D3H2LN cells to Nu/Nu mice, systemic metastases developed in the injected animals. Bioluminescence ...

의학

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 ...

In vivo Dual Substrate Bioluminescent Imaging

Our understanding of how and when breast cancer cells transit from established primary tumors to metastatic sites has increased at an exceptional rate since the advent of in vivo bioluminescent imaging technologies 1-3. Indeed, the ability to locate and quantify tumor growth longitudinally in a single cohort of animals to completion of the study as opposed to sacrificing individual groups of animals at specific assay times has revolutionized how researchers investigate breast cancer metastasis. Unfortunately, current methodologies preclude the real-time assessment of critical changes that transpire in cell signaling systems as breast cancer cells (i) evolve within primary tumors, (ii) disseminate throughout the body, and (iii) reinitiate proliferative programs at sites of a metastatic lesion. However, recent advancements in bioluminescent imaging now make it possible to simultaneously quantify specific spatiotemporal changes in gene expression as a function of tumor development and metastatic progression via the use of dual substrate luminescence reactions. To do so, researchers take advantage for two light-producing luciferase enzymes isolated from the firefly (Photinus pyralis) and sea pansy (Renilla reniformis), both of which react to mutually exclusive substrates that previously facilitated their wide-spread use in in vitro cell-based reporter gene assays 4. Here we demonstrate the in vivo utility of these two enzymes such that one luminescence reaction specifically marks the size and location of a developing tumor, while the second luminescent reaction serves as a means to visualize the activation status of specific signaling systems during distinct stages of tumor and metastasis development. Thus, the objectives of this study are two-fold. First, we will describe the steps necessary to construct dual bioluminescent reporter cell lines, as well as those needed to facilitate their use in visualizing the spatiotemporal regulation of gene expression during specific steps of the metastatic cascade. Using the 4T1 model of breast cancer metastasis, we show that the in vivo activity of a synthetic Smad Binding Element (SBE) promoter was decreased dramatically in pulmonary metastasis as compared to that measured in the primary tumor 4-6. Recently, breast cancer metastasis was shown to be regulated by changes within the primary tumor microenvironment and reactive stroma, including those occurring in fibroblasts and infiltrating immune cells 7-9. Thus, our second objective will be to demonstrate the utility of dual bioluminescent techniques in monitoring the growth and localization of two unique cell populations harbored within a single animal during breast cancer growth and metastasis.

In vivo Dual Substrate Bioluminescent Imaging

Herein we describe the methods to construct, visualize, and quantify the bioluminescent reactions of both firefly and renilla luciferase enzymes expressed in metastatic breast cancer cells during their growth and metastasis in vivo.

생체 듀얼 기판 발광 생물 이미징에

Our understanding of how and when breast cancer cells transit from established primary tumors to metastatic sites has increased at an exceptional rate ...

Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas

The mouse glioma 261 (GL261) is recognized as an in vivo model system that recapitulates many of the features of human glioblastoma multiforme (GBM). The cell line was originally induced by intracranial injection of 3-methyl-cholantrene into a C57BL/6 syngeneic mouse strain 1; therefore, immunologically competent C57BL/6 mice can be used. While we use GL261, the following protocol can be used for the implantation and monitoring of any intracranial mouse tumor model. GL261 cells were engineered to stably express firefly luciferase (GL261-luc). We also created the brighter GL261-luc2 cell line by stable transfection of the luc2 gene expressed from the CMV promoter. C57BL/6-cBrd/cBrd/Cr mice (albino variant of C57BL/6) from the National Cancer Institute, Frederick, MD were used to eliminate the light attenuation caused by black skin and fur. With the use of albino C57BL/6 mice; in vivo imaging using the IVIS Spectrum in vivo imaging system is possible from the day of implantation (Caliper Life Sciences, Hopkinton, MA). The GL261-luc and GL261-luc2 cell lines showed the same in vivo behavior as the parental GL261 cells. Some of the shared histological features present in human GBMs and this mouse model include: tumor necrosis, pseudopalisades, neovascularization, invasion, hypercellularity, and inflammation 1.
Prior to implantation animals were anesthetized by an intraperitoneal injection of ketamine (50 mg/kg), xylazine (5 mg/kg) and buprenorphine (0.05 mg/kg), placed in a stereotactic apparatus and an incision was made with a scalpel over the cranial midline. A burrhole was made 0.1mm posterior to the bregma and 2.3mm to the right of the midline. A needle was inserted to a depth of 3mm and withdrawn 0.4mm to a depth of 2.6mm. Two &mu;l of GL261-luc or GL261-luc2 cells (107 cells/ml) were infused over the course of 3 minutes. The burrhole was closed with bonewax and the incision was sutured.
Following stereotactic implantation the bioluminescent cells are detectable from the day of implantation and the tumor can be analyzed using the 3D image reconstruction feature of the IVIS Spectrum instrument. Animals receive a subcutaneous injection of 150&mu;g luciferin /kg body weight 20 min prior to imaging. Tumor burden is quantified using mean tumor bioluminescence over time. Tumor-bearing mice were observed daily to assess morbidity and were euthanized when one or more of the following symptoms are present: lethargy, failure to ambulate, hunched posture, failure to groom, anorexia resulting in &gt;10% loss of weight. Tumors were evident in all of the animals on necropsy.

Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas

Intracranial implantation of GL261 cells into C57BL/6 mice produces malignant gliomas that recapitulate many of the hallmarks of human glioblastoma multiforme. We used GL261 cells stably expressing luciferase to allow us to use in vivo imaging to follow tumor progression. The surgery and 3D in vivo imaging are demonstrated.

이후 3D로 Intracranial 주입 VIVO에서</emMurine Gliomas의> 발광 생물 이미징

The mouse glioma 261 (GL261) is recognized as an in vivo model system that recapitulates many of the features of human glioblastoma multiforme (GBM). The ...

Bioluminescent Bacterial Imaging In Vivo

This video describes the use of whole body bioluminesce imaging (BLI) for the study of bacterial trafficking in live mice, with an emphasis on the use of bacteria in gene and cell therapy for cancer. Bacteria present an attractive class of vector for cancer therapy, possessing a natural ability to grow preferentially within tumors following systemic administration. Bacteria engineered to express the lux gene cassette permit BLI detection of the bacteria and concurrently tumor sites. The location and levels of bacteria within tumors over time can be readily examined, visualized in two or three dimensions. The method is applicable to a wide range of bacterial species and tumor xenograft types. This article describes the protocol for analysis of bioluminescent bacteria within subcutaneous tumor bearing mice. Visualization of commensal bacteria in the Gastrointestinal tract (GIT) by BLI is also described. This powerful, and cheap, real-time imaging strategy represents an ideal method for the study of bacteria in vivo in the context of cancer research, in particular gene therapy, and infectious disease. This video outlines the procedure for studying lux-tagged E. coli in live mice, demonstrating the spatial and temporal readout achievable utilizing BLI with the IVIS system.

Bioluminescent Bacterial Imaging In Vivo

This article describes the administration of lux-tagged bacteria to mice and subsequent in vivo analysis using IVIS bioluminescence imaging.

발광 생물 세균 이미징<em&gt; 생체 내</em

This video describes the use of whole body bioluminesce imaging (BLI) for the study of bacterial trafficking in live mice, with an emphasis on the use of ...

면역학 및 감염병학

Monitoring Breast Cancer Growth and Metastatic Colony Formation in Mice using Bioluminescence

Breast cancer is a frequent heterogeneous malignancy and the second leading cause of mortality in women, mainly due to distant organ metastasis. Several animal models have been generated, including the widely used orthotopic mouse models, where cancer cells are injected into the mammary fat pad. However, these models cannot help monitor tumor growth kinetics and metastatic colonization. Cutting-edge tools to monitor cancer cells in real time in mice will significantly advance the understanding of tumor biology. 
Here, breast cancer cell lines stably expressing luciferase and green fluorescent protein (GFP) were established. Specifically, this technique contains two sequential steps initiated by measuring the luciferase activity in vitro and followed by the implantation of the cancer cells into mammary fat pads of nonobese diabetic-severe combined immunodeficiency (NOD-SCID) mice. After the injection, both the tumor growth and metastatic colonization are monitored in real time by the noninvasive bioluminescence imaging system. Then, the quantification of GFP-expressing metastases in the lungs will be examined by fluorescence microscopy to validate the observed bioluminescence results. This sophisticated system combining luciferase and fluorescence-based detection tools evaluates cancer metastasis in vivo, which has great potential for use in breast cancer therapeutics and disease management.

Here, we describe a noninvasive monitoring method involving luciferase and green fluorescent protein expression in various breast cancer cell lines. This protocol provides a technique to monitor tumor formation and metastatic colonization in real time in mice.

생체 발광을 사용하여 마우스의 유방암 성장 및 전이성 콜로니 형성 모니터링

Breast cancer is a frequent heterogeneous malignancy and the second leading cause of mortality in women, mainly due to distant organ metastasis. Several ...

암 연구학

In Vivo Imaging to Measure Spontaneous Lung Metastasis of Orthotopically-injected Breast Tumor Cells

Metastasis remains the primary cause of cancer-related death. The succession of events that characterize the metastatic cascade presents multiple opportunities for therapeutic intervention, and the ability to accurately model them in mice is critical to evaluate their effects. Here, a step-by-step protocol is presented for the establishment of orthotopic primary breast tumors and the subsequent monitoring of the establishment and growth of metastatic lesions in the lung using in vivo bioluminescence imaging. This methodology allows for the evaluation of treatment or its biological effects along the entire range of metastatic development, from primary tumor escape to outgrowth in the lungs. Breast orthotopic tumors are generated in mice via injection of a luciferase-labeled cell suspension in the 4th mammary gland. Tumors are allowed to grow and disseminate for a specific amount of time and are then surgically resected. Upon resection, spontaneous lung metastasis is detected, and the growth over time is monitored using in vivo bioluminescence imaging. At the desired experimental endpoint, lung tissue can be collected for downstream analysis. The treatment of established, clinically evident metastasis is critical to improve outcomes for stage IV cancer patients, and it can be evaluated through tail vein models of experimental lung metastasis. However, metastatic dissemination occurs early in breast cancer, and many patients have latent, subclinical disseminated disease after surgery. Utilization of spontaneous models such as this one provides the opportunity to study the whole spectrum of the disease, especially the systemic effects driven by treatment of the primary tumor such as pre-metastatic niche priming, and evaluate treatments on dormant and subclinical disease after surgery.

<meta charset="utf8"></head><body>orthotopically 주입된 유방 종양 세포의 자발적 폐 전이를 측정하기 위한 In vivo imaging

The manuscript describes a methodology for the establishment as well as longitudinal growth monitoring of spontaneous lung metastasis from orthotopically-injected breast tumors, amenable to intervention at all stages of the metastatic cascade.

생체 내 Orthotopically injected breast tumor cells의 자발적 폐 전이를 측정하기 위한 이미징

Metastasis remains the primary cause of cancer-related death. The succession of events that characterize the metastatic cascade presents multiple ...

Combined In vivo Optical and &#181;CT Imaging to Monitor Infection, Inflammation, and Bone Anatomy in an Orthopaedic Implant Infection in Mice

Multimodality imaging has emerged as a common technological approach used in both preclinical and clinical research. Advanced techniques that combine in vivo optical and &#956;CT imaging allow the visualization of biological phenomena in an anatomical context. These imaging modalities may be especially useful to study conditions that impact bone. In particular, orthopaedic implant infections are an important problem in clinical orthopaedic surgery. These infections are difficult to treat because bacterial biofilms form on the foreign surgically implanted materials, leading to persistent inflammation, osteomyelitis and eventual osteolysis of the bone surrounding the implant, which ultimately results in implant loosening and failure. Here, a mouse model of an infected orthopaedic prosthetic implant was used that involved the surgical placement of a Kirschner-wire implant into an intramedullary canal in the femur in such a way that the end of the implant extended into the knee joint. In this model, LysEGFP mice, a mouse strain that has EGFP-fluorescent neutrophils, were employed in conjunction with a bioluminescent Staphylococcus aureus strain, which naturally emits light. The bacteria were inoculated into the knee joints of the mice prior to closing the surgical site. In vivo bioluminescent and fluorescent imaging was used to quantify the bacterial burden and neutrophil inflammatory response, respectively. In addition, &#956;CT imaging was performed on the same mice so that the 3D location of the bioluminescent and fluorescent optical signals could be co-registered with the anatomical &#956;CT images. To quantify the changes in the bone over time, the outer bone volume of the distal femurs were measured at specific time points using a semi-automated contour based segmentation process. Taken together, the combination of in vivo bioluminescent/fluorescent imaging with &#956;CT imaging may be especially useful for the noninvasive monitoring of the infection, inflammatory response and anatomical changes in bone over time.

Combined In vivo Optical and  &#181;CT Imaging to Monitor Infection, Inflammation, and Bone Anatomy in an Orthopaedic Implant Infection in Mice

Combined optical and &#956;CT imaging in a mouse model of orthopaedic implant infection, utilizing a bioluminescent engineered strain of Staphylococcus aureus, provided the capability to noninvasively and longitudinally monitor the dynamics of the bacterial infection, as well as the corresponding inflammatory response and anatomical changes in the bone.

결합<em&gt; 생체</em&gt; 광학 및 μCT 이미징 마우스의 정형 외과 임플란트 감염에 감염, 염증 및 뼈 해부학을 모니터링

Multimodality imaging has emerged as a common technological approach used in both preclinical and clinical research. Advanced techniques that combine in ...

IVIS 스펙트럼을 사용하여 내유 종양의 생체내 이미징에서 발광 생물

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요약

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Bioluminescence 이미징을 사용하여 종양 성장과 치료에 반응 이후 분석 Intracranial 뇌종양의 Xenografts 구축

Bioluminescence 및 마이크로 중부 표준시 이미징과 함께 종양의 Metastases 및 Osteolytic 조직 손상을 모니터링

유방암 뇌 전이의 종양 Hypoxia 역학 생체내의 Bioluminescence 이미징에서는 마우스 모델에서

생체 듀얼 기판 발광 생물 이미징에

이후 3D로 Intracranial 주입 VIVO에서 발광 생물 이미징

발광 생물 세균 이미징

생체 발광을 사용하여 마우스의 유방암 성장 및 전이성 콜로니 형성 모니터링

생체 내 Orthotopically injected breast tumor cells의 자발적 폐 전이를 측정하기 위한 이미징

생체 내 Orthotopically injected breast tumor cells의 자발적 폐 전이를 측정하기 위한 이미징

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