Name: an orthotopic mouse model of spontaneous breast cancer metastasis
Uploaded: 2016-08-14T13:00:00.000+00:00
Duration: 8 min 11 s
Description: Watch this Scientific Journal Video about An Orthotopic Mouse Model of Spontaneous Breast Cancer Metastasis at JoVE.com

Supported by grants from the National Institute of Health grants CA133085, CA182518 and CA185909 (to KL) and VA Merit Review Award BX001962 (to KL).

所有的程序遵循摄政佐治亚大学动物使用及护理委员会指导原则和批准协议。 1.原位乳腺癌肿瘤的建立<ol><li>实验前一天，培养约2×10 6个 4T1-吕克肿瘤细胞在10厘米的培养皿在10ml RPMI培养基中含有10％FBS。孵育在CO 2培养箱中的培养皿，在37℃和5％的CO 2。 </li><li>对肿瘤细胞注射当天，从与真空培养皿除去培养基，加入5毫升无菌磷酸盐缓冲盐水（PBS），pH为7.4至培养皿中，并轻轻摇动培养皿洗培养表面，除去PBS通过真空。 </li><li>加2ml胰蛋白酶-EDTA（0.05％胰蛋白酶和0.53毫摩尔EDTA）的培养皿中，在37℃下孵育其中的CO 2培养箱中约5分钟。检查细胞上倒置显微镜，以确保所有细胞从培养皿底部剥离。&lt;/ LI&gt; <li>通过加入8毫升含有血清的培养基，转移细胞到15毫升锥形管中，并离心将细胞以大约450×g离心在RT 3分钟猝灭胰蛋白酶。 </li><li>在10ml PBS中除去由真空和悬浮细胞上清液。离心细胞以大约450×g离心3分钟。通过真空除去上清液，并重新悬浮在10ml PBS中的细胞。 </li><li>移取10微升的细胞悬液到玻璃罩下血球。计数细胞，以确定细胞浓度。悬浮细胞在PBS中以2×10 5个细胞/ ml的HBSS的密度。转移细胞到1.5ml无菌微量离心管中，并保持细胞在冰上直到使用。 </li><li>使用6只雌性BALB / c小鼠 - 8周为肿瘤细胞注射。 <ol><li>附近刮胡子使用电子头发剪乳头周围3号的头发。 4T1细胞进入乳腺3脂肪垫的注射重复性产生肺转移。用蘸取70％ethano的棉签清洁剃光面积湖3次 - 颠倒的离心管2悬浮细胞。轻轻吸50微升细胞悬液成半CC 27 G 1/2结核菌素注射器。 </li><li>注入下一个BALB / c小鼠的＃3乳腺肿瘤细胞到乳腺脂肪垫。鼠标放置在干净的笼子里，并返回笼子的动物住房设施。等待大约21 - 30天为肿瘤的发展。肿瘤的生长应该定期按机构的对肿瘤的研究策略进行监测。 </li></ol></li></ol> 2.肿瘤生长的活老鼠生物发光成像<ol><li>图像小鼠后每3天肿瘤注射转移小鼠的摄像设备。注入100μl的荧光素（在PBS中30毫克/毫升）腹腔内（IP）给小鼠成像之前。经过大约10分钟，麻醉小鼠用2％异氟醚/ O 2流量感应室。使用镊子去摸鼠标了腿。触摸无反应证实塔T中的鼠标完全不省人事。 <ol><li>放置在连续的2％异氟烷和O 2成像室小鼠以2升/分钟的流速。鼠标放置，使得所感兴趣的区域（ 即 ，初始肿瘤注射的乳腺垫）面向所述成像系统的照相机。确保鼻子和鼠标的嘴的麻醉剂管路内被牢固地设置。动物应每兽药标准在该机构进行麻醉。 </li></ol></li><li>获得生物发光成像的各个曝光时间的阵列。例如，设置曝光至30日，（FOV）视场为25，物体高度为1.5厘米，并获得具有低功耗的X射线荧光照片图像。设置测量单位为"光辉"。 <ol><li>使用自动线性颜色范围内最大约2.4×10 7四射单位。获得初始图像后，调整设置并获得优化的图像（ 图第3和4）。 </li></ol></li><li>回到老鼠笼子清洁，确保老鼠返回动物住房设施之前重新获得下地活动的意识。 </li><li>用与成像仪器相关的生活成像软件分析数据。使用映像程序，画出感兴趣区域（ROI）的区域周围封闭线。使用图像处理软件，以确定投资回报率的光辉力度。记录每个ROI每只小鼠的相对强度。 注:较高的辐射强度指示发光单元的更高水平，并因此更大的肿瘤生长。 </li><li>目视检查小鼠，每3天的不良影响。 注:老鼠会每周一次进行权衡。 15％体重的重量损失被认为是不利的影响，如在步骤5中小鼠溃烂肿瘤也将安乐死描述的荷瘤小鼠进行安乐死。 </li></ol> 3.原发性肿瘤的手术切除没有TE:蒸压剪刀，镊子和伤口夹和伤口施夹器（ 图1）。 <ol><li>在无菌罩下进行手术，以保持无菌状态，以减少感染的风险。手术前一天，剃包围荷瘤小鼠的肿瘤用电动毛发修剪器的区域。 </li><li>约2 - 手术前4小时，进料配制用于小鼠小鼠咀嚼卡布洛芬片剂，剂量为5毫克/公斤体重。 </li><li>把小鼠在含有2％异氟烷在氧气中约30秒的腔室。在整个手术期间使用的麻醉机不断麻醉鼠标。 </li><li>用消毒棉棍兽医软膏适用于鼠标的眼睛，防止干燥时的麻醉下。使用镊子去摸鼠标了腿。触摸没有反应证实了鼠标完全不省人事。 </li><li>消毒肿瘤周围部位的皮肤区域与三个交替抹肥皂擦洗消毒。擦拭用70％的酒精擦拭手术区。 </li><li>用无菌解剖刀（ 图1）切割肿瘤附近的一个小切口。 </li><li>插入一对灭菌剪刀的尖端切口切开皮肤肿瘤周围以暴露肿瘤结节（ 图2A）。持用无菌镊子肿瘤并使用剪刀的肿瘤从皮肤（ 图2B）中分离出来。通过在-80℃冷冻器冷冻或通过在供将来使用10％福尔马林溶液固定保存肿瘤样品。 </li><li>关闭手术部位与使用自动施夹器（ 图1）9毫米卷绕夹子。返回鼠标到一个干净的笼子蒸压床上用品。动物应该每个机构准则进行监控。 注:鼠标通常收复足够的意识，并开始在笼子里大约10移动 - 30分钟后手术。 </li><li>辖卡布洛芬到手术后的小鼠再次24小时。卡洛芬是手术后镇痛和手术后镇痛处方应该由该机构的兽医人员直接协商进行验证。 （ - 手术后7天通常在5）在伤口愈合后伤口取出夹子。除去使用自动剪辑去除缠绕剪辑。 </li></ol> 4.肿瘤转移的活老鼠生物发光成像<ol><li>每隔3天手术后图像小鼠如步骤2所述。 </li></ol> 5.肺转移瘤验证的使用荷瘤肺的墨汁通货膨胀注:要验证荧光素酶活肿瘤​​成像效果，执行荷瘤小鼠肺的墨汁通胀量化肿瘤结节。 4T1细胞也转移到其他器官和组织（ 图5），因此，如果需要的话，应执行这些器官以验证肿瘤转移的组织学检查。该协议使用LUN摹转移作为一个例子。 <ol><li>养老鼠在自己现有的家庭笼安乐死。当安置与其他动物，搬迁不被安乐死到不同的笼子里那些老鼠。 </li><li>移除笼滤波器顶部并与CO 2的过滤器顶部替换。打开被连接到CO 2滤波器顶部的压缩的 CO 2气体筒（100％）。定在2升/分钟的流速，并保持CO 2的上为至少10分钟。如果将小鼠仍然在10分钟结束时呼吸，离开上的 CO 2，直到至少1分钟的小鼠停止呼吸之后。关闭CO 2的气缸控制和流量计。等待3分钟，取出过滤器顶部。 </li><li>放置牺牲鼠标在其上聚苯板回来。针腿以确保对气管畅通无阻。喷用70％乙醇的小鼠。 </li><li>使用剪刀，沿着鼠标的-腹部中间的中线切穿胸腔和向上朝向SalI位有所不同的腺体。观察气管。使用钳去除周围气管暴露气管组织。 </li><li>螺纹气管下枪头。保持尖端用一只手，轻轻向上和从主体向外移动气管。 </li><li>旋转平台上按住鼠标180°。用一个半的CC 27 G 1/2结核菌素注射器注入印度油墨（10％印度墨水和0.1％氢氧化铵）到经由气管肺。完全用墨膨胀肺部，直到强烈感觉到阻力。 </li><li>用剪刀切开气管。使用镊子按住鼠标并插入肺部下另一套镊子和拉肺叶了鼠标。冲洗在含烧杯水肺部短暂。 </li><li>在化学通风橱，肺部转移到装有3毫升的Fekete的溶液（50％乙醇，6％甲醛，和3％冰醋酸）的玻璃闪烁管中。盖上小瓶，以防止从蒸发溶液。 注意:将组织可以存储在菲克特的溶液无限期。 </li><li>几分钟后，观察肿瘤结节为白点的黑色肺（ 图5）。白色结节肿瘤是历历在目。转移肺部菜在通风橱并计数白点的数目。每一个白点代表一个单独的转移性肿瘤结节。 </li></ol>

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Mice were purchased from the Jackson Laboratory and housed in the Georgia Regents University animal facility. Experiments and care/welfare were in agreement with federal regulations and an approved protocol by the Georgia Regents University IACUC committee. The authors have nothing to disclose.

许多类型的乳腺癌转移的转基因小鼠模型已经被开发。1这些转基因小鼠具有高的肿瘤的发病率范围从60到100％。然而，这些转基因小鼠的转移发生率比肿瘤发病率要低得多（14 - 100％）。肿瘤细胞转移至LN和肺中的大多数乳腺癌转移的这些转基因小鼠模型。转移延迟从模型变化以模拟和从2〜8个月的范围内。6,11-16乳腺癌转移的这些转基因小鼠模型可用于研究乳腺癌转移基础的遗传...

根据美国癌症协会，乳腺癌是女性癌症在美国是最常见的诊断形式。在最近开发的靶向疗法组合早期检测已经显著减少乳腺癌的死亡率在过去二十年。然而，乳腺癌仍然是妇女在美国1癌症相关死亡的第二大原因。大多数乳腺癌患者的死亡是由于肿瘤细胞转移。不幸的是，大多数的乳腺癌是浸润性和经常转移至淋巴结，并随后到远处器官，包括骨，肺，肝和脑。1,2-目前转移性乳腺癌没有有效的治疗方法。因此，化疗和免疫治疗剂的发展来抑制转移性乳腺癌具有十分重要的意义。 各种乳腺癌自发转移小鼠模型bEEN开发研究乳腺肿瘤细胞进展和转移，也可以用作模型治疗剂的发展的分子机制。1,3-5然而，大多数这些小鼠模型是遗传肿瘤模型，虽然优良模型机械研究，不适合用于测试治疗剂，因为转移需要几个月在这些遗传模型来开发，因此需要的抗癌剂的昂贵长期施用。在活体小鼠6,7-监测肿瘤进展也是技术上具有挑战性。相反，移植乳腺癌转移模型具有短期肿瘤进展和在活体小鼠的肿瘤进展的容易跟踪的优点。在4T1原位乳腺癌自发转移的小鼠模型是这样的移植肿瘤模型8在这个模型中，乳腺肿瘤细胞移植到乳房脂肪垫建立原发性肿瘤结节。原发性肿瘤然后可在人类乳腺癌患者进行手术切除。 4T1肿瘤细胞是高度侵入性的。8,9，3,10几乎所有携带肿瘤的小鼠发展转移在30天后肿瘤移植到乳房脂肪垫后。然而，4T1肿瘤的原发部位长出积极与肿瘤大小往往超出了允许的大多数动物协议的限制。在这个阶段中，转移往往是微转移。因此，重要的是去除原发肿瘤，以允许用于测试治疗剂转移的进展。这里，我们报告建立原发性肿瘤移植，手术切除原发肿瘤和4T1肿瘤的生长和发展在体内的生物发光基于成像的量化的一个简单且敏感的程序。

<table><tbody><tr><td>Ami-X Imaging System&amp;nbsp;</td><td>Spectral Instruments Imaging Inc. Tucson, AZ</td><td></td><td></td></tr><tr><td>IsoTec SurgiVet</td><td>Anesthesia Service &amp;amp; Equipment , Inc., Atlanta, GA</td><td></td><td></td></tr><tr><td>AutoClip Physicians Kit</td><td>Becton Dickinson Primary Care Diagnostics. Sparks, MD</td><td>427638</td><td></td></tr><tr><td>9 mm AutoClip Applier&amp;nbsp;</td><td>Becton Dickinson Primary Care Diagnostics. Sparks, MD</td><td>427630</td><td></td></tr><tr><td>9 mm AutoClip Remover</td><td>Becton Dickinson Primary Care Diagnostics. Sparks, MD</td><td>427637</td><td></td></tr><tr><td>9 mm AutoClip Wound Clip</td><td>Becton Dickinson Primary Care Diagnostics. Sparks, MD</td><td>427631</td><td></td></tr><tr><td>Sharp-Pointed Dissecting Scissors</td><td>Fisher</td><td>8940</td><td></td></tr><tr><td>Dissecting Fine-Pointed Forceps</td><td>Fisher</td><td>8875</td><td></td></tr><tr><td>1/2 CC 27&amp;nbsp;G&amp;nbsp;1/2 tuberculin syringe</td><td>Becton Dickinson and Co. NJ&amp;nbsp;</td><td>305620</td><td></td></tr><tr><td>RPMI 1640 medium</td><td>Mediatech Inc</td><td>10-040-CV</td><td></td></tr><tr><td>PBS</td><td>Mediatech Inc</td><td>21-040-CV</td><td></td></tr><tr><td>70% Ethanol</td><td>Ultrapure-usa.com</td><td></td><td></td></tr><tr><td>Trypsin-EDTA</td><td>Mediatech Inc</td><td>25-040-CI</td><td></td></tr></tbody></table>

an orthotopic mouse model of spontaneous breast cancer metastasis

转移是乳腺癌患者的死亡的首要原因。该机制癌细胞转移，包括乳腺癌转移底层，在很大程度上是未知的，是在癌症研究的焦点。各种乳腺癌自发转移小鼠模型已经建立。在这里，我们报告的简化程序，建立原位移植乳腺癌原发灶和由此产生的自发转移，模仿人类乳腺癌转移。用生物发光活肿瘤成像相结合，该小鼠模型允许肿瘤生长和进展动力学进行监测和定量。在这种模式下，4T1-吕克乳腺癌细胞的低剂量（1×10 4个细胞），使用结核菌素注射器注入BALB / c小鼠乳腺脂肪垫。小鼠用萤光素注射，并在使用生物发光成像系统的不同时间点成像。当原发肿瘤生长的大小限制为在IACUC批准的协议（APPRoximately 30天），将小鼠在2％异氟烷和氧气的流量恒定麻醉。肿瘤区域用70％乙醇灭菌。肿瘤周围的小鼠皮肤切除以暴露其用一对无菌剪刀除去肿瘤。原发肿瘤的切除延伸的4T-1荷瘤小鼠的存活一个月。小鼠然后重复成像的转移性肿瘤扩散到远处器官。治疗剂可施用在这一点上，以抑制肿瘤的转移。这种模式是在主站点和进展动力学远处器官量化乳腺癌细胞的生长简单而敏感，因此是研究乳腺癌的生长和恶化，并在体内测试抗转移治疗和免疫治疗剂的优秀典范。

原位乳腺癌小鼠模型的建立 4T1是一个积极的乳腺癌细胞系。小至1×10 4个细胞进入乳房脂肪垫的注射可导致建立一个单一的肿瘤结节中注入（ 图2A）的部位。因此，肿瘤模仿人原发性乳房癌。几乎100％的小鼠发展的原位肿瘤。肿瘤大小可以使用数字测径器或通过活肿瘤成像（ 图3）进行定量。肿瘤是使用生物发光成像系?...

Watch this Scientific Journal Video about An Orthotopic Mouse Model of Spontaneous Breast Cancer Metastasis at JoVE.com

An Orthotopic Mouse Model of Spontaneous Breast Cancer Metastasis

原位乳腺癌原发肿瘤模型和手术切除原发肿瘤，延长生命鼠标产生自发转移的描述。肿瘤的生长和发展进行监测和通过萤光素酶荧光成像定量。

自发性乳腺癌转移的原位小鼠模型

Metastasis is the primary cause of mortality of breast cancer patients. The mechanism underlying cancer cell metastasis, including breast cancer ...

an-orthotopic-mouse-model-of-spontaneous-breast-cancer-metastasis

Research

JoVE Journal

Medicine

30.7K 次观看.  Augusta University. 原位乳腺癌原发肿瘤模型和手术切除原发肿瘤，延长生命鼠标产生自发转移的描述。肿瘤的生长和发展进行监测和通过萤光素酶荧光成像定量。 

视频: An Orthotopic Mouse Model of Spontaneous Breast Cancer Metastasis

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.

门静脉注射模型来研究肝转移乳腺癌

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 ...

科研

癌症研究

Orthotopic Injection of Breast Cancer Cells into the Mice Mammary Fat Pad

A proper animal model is crucial for a better understanding of diseases. Animal models established by different methods (subcutaneous injections, xenografts, genetic manipulation, chemical reagents induction, etc.) have various pathological characters and play important roles in investigating certain aspects of diseases. Although no single model can totally mimic the whole human disease progression, orthotopic organs disease models with a proper stromal environment play an irreplaceable role in understanding diseases and screening for potential drugs. In this article, we describe how to implant breast cancer cells into the mammary fat pad in a simple, less invasive, and easy-to-handle way, and follow the metastasis to distant organs. With the proper features of primary tumor growth, breast and nipple pathological changes, and a high occurrence of other organs&#39; metastasis, this model maximumly mimics human breast cancer progression. Primary tumor growth in situ, long-distant metastasis, and the tumor microenvironment of breast cancer can be investigated by using this model.

Here, we present a protocol to implant breast cancer cells into the mammary fat pad in a simple, less invasive, and easy-to-handle way, and this mouse orthotopic breast cancer model with a proper mammary fat pad environment can be used to investigate various aspects of cancer.

乳腺癌细胞原位注射到小鼠乳房脂肪板

A proper animal model is crucial for a better understanding of diseases. Animal models established by different methods (subcutaneous injections, ...

Studying Triple Negative Breast Cancer Using Orthotopic Breast Cancer Model

Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited therapeutic options. When compared to patients with less aggressive breast tumors, the 5-year survival rate of TNBC patients is 77% due to their characteristic drug-resistant phenotype and metastatic burden. Toward this end, murine models have been established aimed at identifying novel therapeutic strategies limiting TNBC tumor growth and metastatic spread. This work describes a practical guide for the TNBC orthotopic model where MDA-MB-231 breast cancer cells suspended in a basement membrane matrix are implanted in the fourth mammary fat pad, which closely mimics the cancer cell behavior in humans. Measurement of tumors by caliper, lung metastasis assessment via in vivo and ex vivo imaging, and molecular detection are discussed. This model provides an excellent platform to study therapeutic efficacy and is especially suitable for the study of the interaction between the primary tumor and distal metastatic sites.

This work presets an advanced protocol to accurately assess tumor loading by detection of green fluorescent protein and bioluminescence signals as well as the integration of quantitative molecular detection technique.

研究使用正交乳腺癌模型的三阴性乳腺癌

Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited therapeutic options. When compared to patients with less ...

Orthotopic Transplantation of Breast Tumors as Preclinical Models for Breast Cancer

Preclinical models that faithfully recapitulate tumor heterogeneity and therapeutic response are critical for translational breast cancer research. Immortalized cell lines are easy to grow and genetically modify to study molecular mechanisms, yet the selective pressure from cell culture often leads to genetic and epigenetic alterations over time. Patient-derived xenograft (PDX) models faithfully recapitulate the heterogeneity and drug response of human breast tumors. PDX models exhibit a relatively short latency after orthotopic transplantation that facilitates the investigation of breast tumor biology and drug response. The transplantable genetically engineered mouse models allow the study of breast tumor immunity. The current protocol describes the method to orthotopically transplant breast tumor fragments into the mammary fat pad followed by drug treatments. These preclinical models provide valuable approaches to investigate breast tumor biology, drug response, biomarker discovery and mechanisms of drug resistance.

Patient-derived xenograft (PDX) models and transplantable genetically engineered mouse models faithfully recapitulate human disease and are preferred models for basic and translational breast cancer research. Here, a method is described to orthotopically transplant breast tumor fragments into the mammary fat pad to study tumor biology and evaluate drug response.

乳腺癌临床前模型乳腺肿瘤的矫形移植

Preclinical models that faithfully recapitulate tumor heterogeneity and therapeutic response are critical for translational breast cancer research. ...

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>体内成像测量原位注射乳腺肿瘤细胞的自发性肺转移

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.

体内研究 影像学检查测量原位注射乳腺肿瘤细胞的自发性肺转移

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

Initiation of Metastatic Breast Carcinoma by Targeting of the Ductal Epithelium with Adenovirus-Cre: A Novel Transgenic Mouse Model of Breast Cancer

Breast cancer is a heterogeneous disease involving complex cellular interactions between the developing tumor and immune system, eventually resulting in exponential tumor growth and metastasis to distal tissues and the collapse of anti-tumor immunity. Many useful animal models exist to study breast cancer, but none completely recapitulate the disease progression that occurs in humans. In order to gain a better understanding of the cellular interactions that result in the formation of latent metastasis and decreased survival, we have generated an inducible transgenic mouse model of YFP-expressing ductal carcinoma that develops after sexual maturity in immune-competent mice and is driven by consistent, endocrine-independent oncogene expression. Activation of YFP, ablation of p53, and expression of an oncogenic form of K-ras was achieved by the delivery of an adenovirus expressing Cre-recombinase into the mammary duct of sexually mature, virgin female mice. Tumors begin to appear 6 weeks after the initiation of oncogenic events. After tumors become apparent, they progress slowly for approximately two weeks before they begin to grow exponentially. After 7-8 weeks post-adenovirus injection, vasculature is observed connecting the tumor mass to distal lymph nodes, with eventual lymphovascular invasion of YFP+ tumor cells to the distal axillary lymph nodes. Infiltrating leukocyte populations are similar to those found in human breast carcinomas, including the presence of &#945;&#946; and &#947;&#948; T cells, macrophages and MDSCs. This unique model will facilitate the study of cellular and immunological mechanisms involved in latent metastasis and dormancy in addition to being useful for designing novel immunotherapeutic interventions to treat invasive breast cancer.

Activation of latent mutations with adenovirus-Cre into the mammary ductal system results in a clinically relevant metastatic breast cancer. Incorporation of a YFP promoter allows tracking of distal metastatic tumor cells. This model is useful to study latent metastasis, anti-tumor immunity, and for designing novel immunotherapies to treat breast cancer.

乳腺癌的一种新的转基因小鼠模型：转移性乳腺癌通过瞄准导管上皮与腺病毒Cre重组酶的启动

Breast cancer is a heterogeneous disease involving complex cellular interactions between the developing tumor and immune system, eventually resulting in ...

医学

Orthotopic Injection of Breast Cancer Cells into the Mammary Fat Pad of Mice to Study Tumor Growth.

Breast cancer growth can be studied in mice using a plethora of models. Genetic manipulation of breast cancer cells may provide insights into the functions of proteins involved in oncogenic progression or help to discover new tumor suppressors. In addition, injecting cancer cells into mice with different genotypes might provide a better understanding of the importance of the stromal compartment. Many models may be useful to investigate certain aspects of disease progression but do not recapitulate the entire cancerous process. In contrast, breast cancer cells engraftment to the mammary fat pad of mice better recapitulates the location of the disease and presence of the proper stromal compartment and therefore better mimics human cancerous disease. In this article, we describe how to implant breast cancer cells into mice orthotopically and explain how to collect tissues to analyse the tumor milieu and metastasis to distant organs. Using this model, many aspects (growth, angiogenesis, and metastasis) of cancer can be investigated simply by providing a proper environment for tumor cells to grow.

Cancer is a complex disease that is influenced by the tissue surrounding the tumor as well as local pro- and anti-inflammatory mediators. Therefore, orthotropic injection models, rather than subcutaneous models may be useful to study cancer progression in a manner that better mimics human pathology.

注射原位乳腺癌细胞到小鼠乳腺脂肪垫来研究肿瘤的生长。

Breast cancer growth can be studied in mice using a plethora of models. Genetic manipulation of breast cancer cells may provide insights into the ...

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.

"以病人为像"肝癌原位同源小鼠模型的癌转移

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 ...

Generating a Murine Orthotopic Metastatic Breast Cancer Model and Performing Murine Radical Mastectomy

In vivo mouse models to assess breast cancer progression are essential for cancer research, including preclinical drug developments. However, the majority of the practical and technical details are commonly omitted in published manuscripts which, therefore, makes it challenging to reproduce the models, particularly when it involves surgical techniques. Bioluminescence technology allows for the evaluation of small amounts of cancer cells even when a tumor is not palpable. Utilizing luciferase-expressing cancer cells, we establish a breast cancer orthotopic inoculation technique with a high tumorigenesis rate. Lung metastasis is assessed utilizing an ex vivo technique. We, then, establish a mastectomy model with a low local recurrence rate to assess the metastatic tumor burden. Herein, we describe, in detail, the surgical techniques of orthotopic implantation and mastectomy for breast cancer with a high tumorigenesis rate and low local recurrence rates, respectively, to improve breast cancer model efficiency.

We introduce a murine orthotopic breast cancer model and radical mastectomy model with bioluminescence technology to quantify the tumor burden to mimic human breast cancer progression.

小鼠原位转移性乳腺癌模型的建立及小鼠根治性乳房切除术

In vivo mouse models to assess breast cancer progression are essential for cancer research, including preclinical drug developments. However, the majority ...

自发性乳腺癌转移的原位小鼠模型

摘要

探索更多视频

此视频中的章节

门静脉注射模型来研究肝转移乳腺癌

乳腺癌细胞原位注射到小鼠乳房脂肪板

研究使用正交乳腺癌模型的三阴性乳腺癌

乳腺癌临床前模型乳腺肿瘤的矫形移植

体内研究影像学检查测量原位注射乳腺肿瘤细胞的自发性肺转移

乳腺癌的一种新的转基因小鼠模型：转移性乳腺癌通过瞄准导管上皮与腺病毒Cre重组酶的启动

体内研究影像学检查测量原位注射乳腺肿瘤细胞的自发性肺转移

注射原位乳腺癌细胞到小鼠乳腺脂肪垫来研究肿瘤的生长。

"以病人为像"肝癌原位同源小鼠模型的癌转移

小鼠原位转移性乳腺癌模型的建立及小鼠根治性乳房切除术

自发性乳腺癌转移的原位小鼠模型

摘要

探索更多视频

此视频中的章节

门静脉注射模型来研究肝转移乳腺癌

乳腺癌细胞原位注射到小鼠乳房脂肪板

研究使用正交乳腺癌模型的三阴性乳腺癌

乳腺癌临床前模型乳腺肿瘤的矫形移植

体内研究 影像学检查测量原位注射乳腺肿瘤细胞的自发性肺转移

乳腺癌的一种新的转基因小鼠模型：转移性乳腺癌通过瞄准导管上皮与腺病毒Cre重组酶的启动

体内研究 影像学检查测量原位注射乳腺肿瘤细胞的自发性肺转移

注射原位乳腺癌细胞到小鼠乳腺脂肪垫来研究肿瘤的生长。

"以病人为像"肝癌原位同源小鼠模型的癌转移

小鼠原位转移性乳腺癌模型的建立及小鼠根治性乳房切除术

体内研究影像学检查测量原位注射乳腺肿瘤细胞的自发性肺转移

体内研究影像学检查测量原位注射乳腺肿瘤细胞的自发性肺转移