Name: mammary transplantation of stromal cells and carcinoma cells in c57bl/6j mice
Uploaded: 2011-08-12T12:00:00
Description: Watch this Scientific Journal Video about Mammary Transplantation of Stromal Cells and Carcinoma Cells in C57BL/6J Mice at JoVE.com

资助这个项目是通过NIH / NCI的授予R00 CA127357和堪萨斯大学癌症研究中心捐赠。

1。 C57BL/6J小鼠捐助胶原蛋白的分离和提取<ol><li>牺牲成熟的正常使用批准IACUC方法雌性C57BL/6J小鼠。 </li><li>收获的尾巴，和70％的乙醇浸泡45分钟消毒组织。干用纸巾的尾巴，在铝箔包装。存放在-20℃的尾巴，直到需要。 </li><li>放置在无菌的环境，如层流罩尾巴。用剪刀，分裂尾根皮肤和剥离从尾部。删除从尾部两端的0.5〜1厘米，并分为三，四件，剩下的尾巴。剖析尾巴筋和分离成单纤维筋，?...

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功能的成纤维细胞在肿瘤进展的贡献已经证明，通过移植模型，癌相关成纤维细胞联合移植良性乳腺上皮细胞，在肿瘤的生长和增加侵袭5结果。涉及传统的移植方法有免疫缺陷或裸鼠移植合作，从不同的遗传鼠标背景或不同物种的基质和上皮细胞的使用。免疫功能低下小鼠缺乏功能性T细胞，从而起到关键作用，通过识别肿瘤特异性抗原和随后的定位，抑制肿瘤细胞转移扩散 6，?...

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>目录编号</th><th>评论（可选） </th></tr><tr><td> C57BL/6N小鼠</td><td>哈伦</td><td> N / A </td><td></td></tr><tr><td> MMTV - PyVmT转基因小鼠</td><td>杰克逊实验室</td><td> 002374 </td><td></td></tr><tr><td>胎牛血清</td><td>费舍尔</td><td> SH3039603PR </td><td></td></tr><tr><td> DMEM培养液</td><td> VWR </td><td> 10000113873 </td><td></td></tr><tr><td>青霉素/链霉素</td><td>费舍尔</td><td> MT - 30 - 001 </td><td></td></tr><tr><td>两性霉素</td><td>费舍尔</td><td> BP2645 - 20 </td><td></td></tr><tr><td> Amicon过滤列ultracel 50K </td><td> Millipore公司</td><td> UFC905008 </td><td></td></tr><tr><td>贝克曼德州仪器转子管</td><td>贝克曼</td><td> 355618 </td><td></td></tr><tr><td>鼠尾胶原</td><td>费舍尔</td><td> CB 40236 </td><td></td></tr><tr><td> 10X EBSS </td><td>西格玛爱秩序</td><td> E7510 - 100ML </td><td></td></tr><tr><td>胰蛋白酶1X的HBSS W / O钙和镁的0.25％ </td><td>费舍尔</td><td> MT - 25 - 050 - CI </td><td></td></tr><tr><td>冰醋酸</td><td>费舍尔</td><td> A491 - 212 </td><td></td></tr><tr><td> Coomasie蓝色</td><td>费舍尔</td><td> BP101 25 </td><td></td></tr><tr><td>胰蛋白酶</td><td>西格玛爱秩序</td><td> T3924 - 100ML </td><td></td></tr><tr><td>胶原酶一个</td><td>适马AldrichC0130 - 50 </td><td></td><td></td></tr><tr><td> hyalronidase </td><td>西格玛爱秩序</td><td> H3884 </td><td></td></tr><tr><td> DNA酶</td><td>西格玛爱秩序</td><td> D5025 </td><td></td></tr><tr><td>万花筒蛋白标准</td><td> BIORAD </td><td> 1610375 </td><td></td></tr><tr><td>载玻片</td><td>费舍尔</td><td> 12545-78 </td><td></td></tr><tr><td>盖玻片</td><td> VWR </td><td> 101400-042 </td><td></td></tr><tr><td>波形蛋白抗体的S - 20 </td><td>圣克鲁斯生物技术</td><td> SC - 7558 </td><td></td></tr><tr><td> α-平滑肌肌动蛋白抗体</td><td> Abcam公司</td><td> ab5694 </td><td></td></tr><tr><td> CK14抗体</td><td>圣克鲁斯生物技术</td><td> SC - 53253 </td><td></td></tr><tr><td> CK18的抗体</td><td> Abcam公司</td><td> ab668 </td><td></td></tr><tr><td> DAPI </td><td>西格玛爱秩序</td><td> D9542 </td><td></td></tr><tr><td>反鼠标生物素</td><td>媒介实验室</td><td> BA9200 </td><td>分布通过费舍尔</td></tr><tr><td>抗鼠ALEXA - 568 </td><td> Invitrogen公司</td><td> A10037 </td><td></td></tr><tr><td>抗鼠ALEXA - 488 </td><td> Invitrogen公司</td><td> A11001 </td><td></td></tr><tr><td>链霉亲和ALEXA - 488 </td><td> Invitrogen公司</td><td> S11226 </td><td></td></tr><tr><td> DAPI </td><td> Invitrogen公司</td><td> D21490 </td><td></td></tr><tr><td>延长抗淬灭</td><td> Invitrogen公司</td><td> P - 36930 </td><td></td></tr><tr><td>手术剪</td><td>精细科学的工具</td><td> 91400-12 </td><td></td></tr><tr><td>鸟语花香，剪刀</td><td>精细科学的工具</td><td> 15000-02 </td><td></td></tr><tr><td>钝钳</td><td>精细科学的工具</td><td> 11002-12 </td><td></td></tr><tr><td> ＃5细钳</td><td>精细科学的工具</td><td> 11251-10 </td><td></td></tr><tr><td>肠道铬缝合</td><td>费舍尔</td><td> NC9326254 </td><td></td></tr><tr><td>玻璃巴斯德吸管</td><td>费舍尔</td><td> 22-042-815 </td><td></td></tr><tr><td>乙醇</td><td>费舍尔</td><td> A406P 4 </td><td></td></tr><tr><td>优碘</td><td>费舍尔</td><td> NC9386574 </td><td></td></tr><tr><td>伤口剪辑</td><td>费舍尔</td><td> 12032-07 </td><td></td></tr><tr><td>伤口主食</td><td>费舍尔</td><td> 12031-07 </td><td></td></tr></tbody></table>

mammary transplantation of stromal cells and carcinoma cells in c57bl/6j mice

基质细胞的影响，包括对乳腺肿瘤的进展的成纤维细胞已通过使用小鼠模型，特别是通过在小鼠乳腺间质细胞和上皮细胞移植。目前的移植模型往往涉及由于不同遗传背景的基质细胞和上皮细胞的免疫功能低下小鼠的使用。细胞外基质经常被用来嵌入一致的细胞 - 细胞相互作用的两种不同类型的细胞，但涉及的基质胶或鼠尾胶原蛋白，它具有免疫原性的基板使用。缺乏功能的T细胞免疫功能低下小鼠防止基质细胞在体内对乳腺肿瘤的进展进行准确的评估，对药物研发和疗效有重要的影响。此外，免疫功能低下小鼠，成本高昂，难以滋生和需要特殊照顾的条件。为了克服这些障碍，我们已经开发出一种方法原位基质细胞和上皮细胞相同的遗传背景移植到小鼠体内诱发肿瘤形成一致。该系统涉及捐助C57BL/6J小鼠收获正常，癌相关成纤维细胞，PyVmT乳腺癌细胞和胶原蛋白。胶原蛋白细胞，然后嵌入和雌性C57BL/6J小鼠腹股沟乳腺移植。单靠PyVmT细胞移植的形式30-40天可扪及肿瘤移植后。端点在60天的分析表明，与成纤维细胞联合移植相比提高了乳腺肿瘤的生长PyVmT细胞移植单独。虽然在这些研究中，C57BL/6J小鼠的细胞和基质细胞和基质隔离和移植的方法可应用于对来自不同遗传背景的小鼠表现出多功能性。综上所述，本系统可用于研究基质细胞和上皮细胞之间的分子间的相互作用，并克服在免疫功能低下小鼠模型中的关键限制。

Watch this Scientific Journal Video about Mammary Transplantation of Stromal Cells and Carcinoma Cells in C57BL/6J Mice at JoVE.com

Mammary Transplantation of Stromal Cells and Carcinoma Cells in C57BL/6J Mice

在这份报告中，我们展示了一个系统，以隔离和文化的供体细胞，从小鼠乳腺和原位移植受体小鼠，这些细胞分析基质上皮细胞的相互作用：在乳腺肿瘤的发展。

C57BL/6J小鼠乳腺间质细胞和癌细胞的移植

The influence of stromal cells, including fibroblasts on mammary tumor progression has been well documented through the use of mouse models, in particular ...

Research

JoVE Journal

Medicine

Experimental Generation of Carcinoma-Associated Fibroblasts (CAFs) from Human Mammary Fibroblasts

Experimental Generation of Carcinoma-Associated Fibroblasts CAFs from Human Mammary Fibroblasts

从人类乳腺成纤维细胞癌相关的成纤维细胞的实验发电（CAFS）

Carcinomas are complex tissues comprised of neoplastic cells and a non-cancerous compartment referred to as the 'stroma'. The stroma consists of ...

科研

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Human Internal Mammary Artery (IMA) Transplantation and Stenting: A Human Model to Study the Development of In-Stent Restenosis

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人乳内动脉（IMA）移植及内支架置入术：人体模型支架内再狭窄的研究开发

Preclinical in vivo research models to investigate pathobiological and pathophysiological processes in the development of intimal hyperplasia after vessel ...

Isolation, Characterization and Comparative Differentiation of Human Dental Pulp Stem Cells Derived from Permanent Teeth by Using Two Different Methods

人牙髓干细胞的分离，鉴定和比较性恒牙使用两种不同的方法，得出

Developing wisdom teeth are easy-accessible source of stem cells during the adulthood which could be obtained by routine orthodontic treatments. Human ...

Generation of Subcutaneous and Intrahepatic Human Hepatocellular Carcinoma Xenografts in Immunodeficient Mice

皮下及肝内人肝癌裸鼠移植瘤的免疫缺陷小鼠的产生

In  vivo experimental models of hepatocellular carcinoma (HCC) that recapitulate  the human disease provide a valuable platform for research into disease  ...

Coculture System with an Organotypic Brain Slice and 3D Spheroid of Carcinoma Cells

与器官型脑片和癌细胞的3D球体共培养系统

Patients with cerebral metastasis of carcinomas have a poor prognosis. However, the process at the metastatic site has barely been investigated, in ...

Modeling Spontaneous Metastatic Renal Cell Carcinoma (mRCC) in Mice Following Nephrectomy

Modeling Spontaneous Metastatic Renal Cell Carcinoma mRCC in Mice Following Nephrectomy

在小鼠模型自发转移性肾细胞癌（MRCC）肾切除

One of the key challenges to improved testing of new experimental therapeutics in renal cell carcinoma (RCC) is the development of models that faithfully ...

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

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 ...

Evaluation of Stem Cell Properties in Human Ovarian Carcinoma Cells Using Multi and Single Cell-based Spheres Assays

评价的人卵巢癌细胞干细胞特性使用多和单细胞为基础的化验球

Years of research indicates that ovarian cancers harbor a heterogeneous mixture of cells including a subpopulation of so-called &#8220;cancer stem ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

研究胰腺癌干细胞特性开发新的治疗策略

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

Heterotopic Cervical Heart Transplantation in Mice

颈椎异位心脏移植小鼠

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies ...