Name: mame models for 4d live-cell imaging of tumor: microenvironment interactions that impact malignant progression
Uploaded: 2012-02-17T12:00:00
Description: Watch this Scientific Journal Video about MAME Models for 4D Live-cell Imaging of Tumor: Microenvironment Interactions that Impact Malignant Progression at JoVE.com

这项工作的支持，在某种程度上，由国家卫生部以R01 CA131990（矿渣和RRM）研究院。活细胞成像显微成像和流式细胞仪的资源支持核心部分，由NIH中心授予P30 CA22453 Karmanos癌症研究所，美国韦恩州立大学和国家儿童健康与发展研究院围产期研究科韦恩州立大学。

1。准备DQ-基板<ol><li>允许冻干DQ-基板预热至室温，然后再打开小瓶，准备去离子水1毫克/毫升DQ的基板库存的解决方案，在4°C到50μL分装和存储鸿沟</li></ol> 这可能是必要的，鼓动〜5分钟，热到50°C，以促进分散的超声波水浴DQ-基板 。 <ol start="2"><li>在冰上解冻一夜之间在4的RBM℃; RBM，应处理在所有时间上冰。 </li><li>使10X磷酸盐缓冲液（PBS），溶解80克氯化钠（1.37男）...

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	<li>Sameni, M., Dosescu, J., Sloane, B. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+imaging+of+proteolysis:+stromal+and+inflammatory+cells+increase+tumor+proteolysis.">Functional imaging of proteolysis: stromal and inflammatory cells increase tumor proteolysis.</a> Mol. Imaging. 2, 159-175 (2003).</li><li>Jedeszko, C., Sameni, M., Olive, M. B., Moin, K., Sloane, B. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visualizing+protease+activity+in+living+cells:+from+two+dimensions+to+four+dimensions.">Visualizing protease activity in living cells: from two dimensions to four dimensions.</a> Curr. Protoc. Cell Biol. 39, (2008).</li><li>Jedeszko, C., Victor, B. C., Podgorski, I., Sloane, B. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fibroblast+hepatocyte+growth+factor+promotes+invasion+of+human+mammary+ductal+carcinoma+in+situ.">Fibroblast hepatocyte growth factor promotes invasion of human mammary ductal carcinoma in situ.</a> Cancer Res. 69, 9148-9155 (2009).</li><li>Sameni, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+live-cell+imaging+demonstrates+that+beta1-integrin+promotes+type+IV+collagen+degradation+by+breast+and+prostate+cancer+cells.">Functional live-cell imaging demonstrates that beta1-integrin promotes type IV collagen degradation by breast and prostate cancer cells.</a> Mol. Imaging. 7, 199-213 (2008).</li><li>Li, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=p21-activated+kinase+1+coordinates+aberrant+cell+survival+and+pericellular+proteolysis+in+a+three-dimensional+culture+model+for+premalignant+progression+of+human+breast+cancer.">p21-activated kinase 1 coordinates aberrant cell survival and pericellular proteolysis in a three-dimensional culture model for premalignant progression of human breast cancer.</a> Neoplasia. 10, 314-328 (2008).</li><li>Sameni, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+and+quantifying+the+dynamics+of+tumor+associated+proteolysis.">Imaging and quantifying the dynamics of tumor associated proteolysis.</a> Clin. Exp. Metastasis. 26, 299-309 (2009).</li><li>Cavallo-Medved, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Live-cell+imaging+demonstrates+that+proteases+in+caveolae+of+endothelial+cells+degrade+extracellular+matrix.">Live-cell imaging demonstrates that proteases in caveolae of endothelial cells degrade extracellular matrix.</a> Exp. Cell. Res. 315, 1234-1246 (2009).</li><li>Fukumura, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+microvasculature+and+microenvironment:+novel+insights+through+intravital+imaging+in+pre-clinical+models.">Tumor microvasculature and microenvironment: novel insights through intravital imaging in pre-clinical models.</a> Microcirculation. 17, 206-225 (2010).</li></ol>

作为证明，MAME的共同培养​​，可用于活细胞成像在乳腺肿瘤和其微环境，包括各种细胞成分之间的相互作用的实时。在我们的实验室正在进行的研究中，我们已经使用MAME的共同培养​​，以确定过渡，从原位癌到浸润性导管癌的蛋白水解途径，以及相关的蛋白水解途径和其他途径参与这种过渡，如趋化因子/细胞因子/生长因子途径之间的相互作用。我们进一步表明MAME的模型可以用来作为筛选?...

<table border="1"><tbody><tr><td> 试剂/设备 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>重组基底膜（Cultrex的） </td><td> trevigen </td><td> 3445-005-01 </td><td>媲美基底膜（BD公司） </td></tr><tr><td> I型胶原</td><td>凝聚力实验室</td><td> 5005-B的</td><td></td></tr><tr><td> DQ-基板 （I型胶原，Ⅳ） </td><td> Invitrogen公司</td><td> DQ-COL-D12060 DQ-COL四D12052 </td><td></td></tr><tr><td> 22毫米的塑料盖玻片</td><td>尔科技有限公司</td><td> 12-547 </td><td>减少了一半，70％的乙醇和空气干燥，使用前10分钟离开。 </td></tr><tr><td>细胞培养基</td><td>龙沙</td><td> MEBM-PRF CC-3153 MEGM的CC-4136 </td><td>无酚红</td></tr><tr><td> ibiTreatμ-碟</td><td> ibidi </td><td> 80136 </td><td></td></tr><tr><td> volocity软件</td><td>在Perkin-Elmer </td><td>版本5.5 </td><td>其他品牌的成像软件，可以用来产生三维重建和电影。 </td></tr></tbody></table>

mame models for 4d live-cell imaging of tumor: microenvironment interactions that impact malignant progression

我们已经开发出三维共培养模型，我们长期MAME的（M ammary 1 rchitecture 和 m icroenvironmenténgineering），用于活细胞成像中实时细胞：细胞间的相互作用。我们的总体目标是发展架构，以原位癌的乳腺病变的研究进展侵入型的模型，概括。具体来说，我们开发的模型来分析预恶性乳腺上皮细胞变异和肿瘤微环境，提高或降低原位癌乳房上皮细胞的浸润性导管癌的进展已牵连的其他类型的细胞之间的相互作用。最新研究其他细胞类型的肌上皮细胞，成纤维细胞，巨噬细胞和血液和淋巴微血管内皮细胞。除了MAME的车型，其目的是概括乳房内的细胞相互作用过程中癌症的发展，我们已经制定了前列腺癌的进展同类机型。 在这里，我们说明建立3D共培养与使用的活细胞成像和功能的蛋白质检测，遵循共同培养的乳腺导管原位癌 （DCIS）的细胞和成纤维细胞，随着时间的推移，侵入型的过渡，在程序这种情况下超过二十三天在文化。 MAME的共同培养​​由多个层组成。成纤维细胞中嵌入底层的I型胶原。放在一层重组基底膜（RBM），其中原位癌细胞接种。最后的2％RBM包括顶层和补充每一个媒体的变化。图像与侵入型的进展相关的蛋白质，我们用染料荧光淬火（DQ）的基质蛋白（DQ胶原我与I型胶原和DQ的IV型胶原混合层与中间层的RB混合M）和观察使用共聚焦显微镜的活文化。光学部分被捕获，处理和Volocity可视化软件的三维重建。在MAME的共同培养​​23天的过程中，原位癌细胞增殖和凝聚到大的侵入结构。成纤维细胞迁移，并成为纳入到这些入侵结构。 DCIS的结构表面的胶原蛋白的荧光蛋白片段协会发现，细胞内，也分散在周围的矩阵。目标蛋白，趋化因子/细胞因子和蛋白激酶途径或修改的共同培养​​的细胞组成，可以减少侵袭，MAME的模型可以用来作为新的治疗方法的临床屏幕的药物。

Watch this Scientific Journal Video about MAME Models for 4D Live-cell Imaging of Tumor: Microenvironment Interactions that Impact Malignant Progression at JoVE.com

MAME Models for 4D Live-cell Imaging of Tumor: Microenvironment Interactions that Impact Malignant Progression

我们已经制定了活细胞成像在乳腺肿瘤细胞和其他细胞之间的相互作用，在自己的微实时三维共培养模型，侵入型的影响进展。这些模型可以作为目标旁诱导蛋白，趋化因子/细胞因子和蛋白激酶途径侵袭牵连的药物临床前屏幕。

MAME的模型四维活细胞成像肿瘤微环境的相互作用，影响的恶性进展

We have developed 3D coculture models, which we term MAME (mammary architecture and microenvironment engineering), and used them for live-cell imaging in ...

Research

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Epithelial ovarian cancers (EOCs) are the leading cause of death from gynecological malignancy in Western societies. Despite advances in surgical ...

Live Imaging of Drug Responses in the Tumor Microenvironment in Mouse Models of Breast Cancer

实时成像在乳腺癌小鼠模型中的肿瘤微环境对药物的反应

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Although bioluminescence imaging (BLI) shows promise for monitoring tumor burden in animal models of cancer, these analyses remain mostly qualitative. ...