Name: isolation of mammary epithelial cells from three-dimensional mixed-cell spheroid co-culture
Uploaded: 2012-04-30T12:00:00
Description: Watch this Scientific Journal Video about Isolation of Mammary Epithelial Cells from Three-dimensional Mixed-cell Spheroid Co-culture at JoVE.com

1。建立三维共培养<ol><li>建立联合培养的前一天，从4℃冰箱中冷冻和解冻冰基底膜一夜之间。 </li><li>在当天的实验，准备共同的文化，它对应的上皮细胞，可用于介质（在这个例子中的三菱电梯中等介质：DME/F12介质与10％小牛血清，5微克/毫升胰岛素1微克/毫升氢化可的松，1毫微克/毫升表皮生长因子）。保持冰至少1小时前与基底膜混合介质。 </li><li>冲淡了解冻的基底膜，确定总的基底膜所需...

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这里介绍的方法代表着一个简单的方法分析，包括肿瘤细胞的上皮细胞间质成纤维细胞的影响。它允许直接的细胞内细胞外基底膜基质的三维背景的互动，作进一步的分析，同时使基质细胞容易提取。这可以应用于研究肿瘤相关基质，成纤维细胞系可以被操纵，影响信号通路的选择和上皮线，可以在不同的侵入性的潜力，如在图1所示。三维共培养模型，使细胞侵袭的变化已经引起视?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>屋宇署基底膜</td><td> BD公司</td><td> 356237 </td><td>酚红免费</td></tr><tr><td>胎牛血清古典液体培养基：DMEM培养基F12键</td><td> ThermoScientific </td><td> SH30023.01 </td><td></td></tr><tr><td>小牛血清胎牛血清牛</td><td> ThermoScientific </td><td> SH3007303 </td><td></td></tr><tr><td>胰岛素</td><td>电解二氧化锰化学品</td><td> 407709 </td><td>溶于盐酸0.005N; 0.22微米过滤器</td></tr><tr><td>氢化可的松</td><td> Sigma-Aldrich公司</td><td> H4001 </td><td>溶于50％乙醇; 0.22微米过滤器</td></tr><tr><td>表皮生长因子</td><td> Sigma-Aldrich公司</td><td> E9644 </td><td> 10mm的醋酸中溶解; 0.22微米过滤器</td></tr></tbody></table>

isolation of mammary epithelial cells from three-dimensional mixed-cell spheroid co-culture

虽然已经进入了巨大的努力识别信号通路和分子参与正常细胞和恶性行为1-2，使用传统的二维细胞培养模型，这项工作已完成，允许细胞操纵方便。它已成为明确的细胞内信号通路的影响外的力量，包括维和细胞的表面张力3-4。已采取多种途径，开发三维模型，更准确地反映生物组织架构3。虽然这些模型纳入多维度和建筑讲，由此产生的对细胞影响的研究是比在两维模型的局限性和困难，在随后的分析中提取细胞组织培养的简便。 重要作用，肿瘤周围的微环境在肿瘤发生的行为我之所以成为越来越认识4。肿瘤基质是由多种细胞类型和细胞外分子。在肿瘤的发展有肿瘤细胞和基质细胞之间的双向信号。虽然参与肿瘤基质协同进化的一些因素已经确定，仍然有必要制定简单的技术，系统地识别和研究这些信号6阵列。成纤维细胞是最丰富的细胞类型，在正常或肿瘤相关间质组织，促进基底膜和旁分泌生长因子7的沉积和维护。 许多团体已经使用三维培养系统研究成纤维细胞的各种细胞功能的作用，包括肿瘤反应治疗，免疫细胞招聘，信号分子，细胞增殖，凋亡，血管生成，侵袭8-15。我们已经优化了屁股的简单方法对乳腺上皮细胞，使用市售的细胞外基质的模型来创建三维混合细胞群（联合培养）16-22文化essing乳腺成纤维细胞的影响。续共培养细胞的形成与球体内部的聚类成纤维细胞和上皮细胞很大程度上取决于外部的球体，形成多细胞的预测，到矩阵。操纵改变上皮细胞的侵袭，导致成纤维细胞可以很容易量化的数量和长度23上皮预测的变化。此外，我们已经制定为隔离上皮细胞三维共培养，促进成纤维细胞的对上皮行为暴露的影响分析的方法。我们已经找到了合作文化的影响，持续数周后上皮细胞的分离，允许充裕的时间来执行多个检测。这种方法是适应细胞不同恶性潜能，不需要专门的设备。这种技术允许多个条件下，在体外细胞模型的快速评估和相应的结果，可以在动物体内组织模型以及人体组织样本。

Watch this Scientific Journal Video about Isolation of Mammary Epithelial Cells from Three-dimensional Mixed-cell Spheroid Co-culture at JoVE.com

Isolation of Mammary Epithelial Cells from Three-dimensional Mixed-cell Spheroid Co-culture

被形容为一个简单的方法，分析相关的上皮细胞组织成纤维细胞的影响。这种方法和三维组织培养相结​​合，可以方便从3D隔离后的细胞分析。该技术适用于不同恶性潜能的细胞，使系统的研究对肿瘤细胞的肿瘤相关基质的影响。

从三维混合细胞球体共培养的乳腺上皮细胞的隔离

While enormous efforts have gone into identifying signaling pathways and molecules involved in normal and malignant cell behaviors1-2, much of this work ...

Research

JoVE Journal

Biology

Mouse Mammary Epithelial Cells form Mammospheres During Lactogenic Differentiation

小鼠乳腺上皮细胞形成过程中Lactogenic分化的微球体

A phenotypic measure commonly used to determine the degree of lactogenic differentiation in mouse mammary epithelial cell cultures is the formation of ...

科研

生物学

Microfluidic Co-culture of Epithelial Cells and Bacteria for Investigating Soluble Signal-mediated Interactions

微流体调查可溶性信号介导的相互作用的上皮细胞和细菌的共同文化

The human gastrointestinal (GI) tract is a unique environment in which intestinal epithelial cells and non-pathogenic (commensal) bacteria coexist. It has ...

Mammary Epithelial Transplant Procedure

乳腺上皮移植手术

This article describes and compares the fat pad clearance procedure developed by DeOme KB et al.1 and the sparing procedure developed by Brill B et al.2, ...

Isolation and Culture of Adult Epithelial Stem Cells from Human Skin

成人从人体皮肤的上皮干细胞的分离和培养

The homeostasis of all self-renewing tissues is dependent on adult stem cells. As undifferentiated stem cells undergo asymmetric divisions, they generate ...

Chromatin Isolation by RNA Purification (ChIRP)

Chromatin Isolation by RNA Purification ChIRP

RNA纯化的染色质隔离（CHIRP）

Long noncoding RNAs are key regulators of chromatin states for important biological processes such as dosage compensation, imprinting, and developmental ...

An Analytical Tool that Quantifies Cellular Morphology Changes from Three-dimensional Fluorescence Images

细胞形态变化，从三维荧光图像量化的分析工具，

The most common software analysis tools available for measuring fluorescence images are for two-dimensional (2D) data that rely on manual settings for ...

Induction and Analysis of Epithelial to Mesenchymal Transition

诱导上皮和分析间质转化

Epithelial  to mesenchymal transition (EMT) is essential for proper morphogenesis during  development. Misregulation of this  process has been implicated ...

Isolation and Culture of Dental Epithelial Stem Cells from the Adult Mouse Incisor

从成年小鼠切牙牙源性上皮干细胞的分离和培养

Understanding the cellular and molecular mechanisms that underlie tooth regeneration and renewal has become a topic of great interest1-4, and the mouse ...

FRET Imaging in Three-dimensional Hydrogels

FRET成像的三维水凝胶

Imaging of F&#246;rster resonance energy transfer (FRET)&#160;is a powerful tool for examining cell biology in real-time. Studies utilizing FRET commonly ...

Evaluating the Angiogenetic Properties of Ovarian Cancer Stem-Like Cells using the Three-Dimensional Co-Culture System, NICO-1

使用三维共培养系统NICO-1评估卵巢癌干细胞样细胞的血管遗传特性

Cancer stem cells (CSCs) reside in a supportive niche, constituting a microenvironment comprised of adjacent stromal cells, vessels, and extracellular ...