Name: three-dimensional cell culture model for measuring the effects of interstitial fluid flow on tumor cell invasion
Uploaded: 2012-07-25T12:00:00
Description: Watch this Scientific Journal Video about Three-dimensional Cell Culture Model for Measuring the Effects of Interstitial Fluid Flow on Tumor Cell Invasion at JoVE.com

1。含量的设置<ol><li>冰在4°C（约2小时），解冻的基底膜小等分（&lt;500μL）。 </li><li>准备凝胶配方（看到表中的例子卷以下）：10倍PBS（总体积的1倍），1N钠氢（相当于0.023补充胶原蛋白的量，或每的胶原蛋白制造商的建议，如适用），基底膜和胶原键入我来终浓度为1毫克/毫升和1.3毫克/毫升（可用于其他基质配方，根据细胞类型和实验）。 </li></ol> 例如凝胶配方...

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在这里，我们所描述的量化对肿瘤细胞浸润间流动的影响，使用一个3-D矩阵嵌入细胞内的细胞培养插入方法。这和其他类似的方法已被用来研究多种细胞类型，13日至15日，17间流动的影响。我们的做法，部分模仿使用类型的肿瘤基质微环境，我胶原和基底膜含有蛋白质发现在基底膜上皮组织和周围基质21-22。这个系统是相对容易的设置了，简单，更具成本比大多数微流体装置，用?...

<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>胶原蛋白（鼠尾） </td><td>屋宇署</td><td> 354236 </td><td>保持无菌</td></tr><tr><td> Millicell小细胞培养插入</td><td> Millipore公司</td><td> PI8P01250 </td><td> 8微米孔径，聚碳酸酯膜</td></tr><tr><td>基底膜</td><td>屋宇署</td><td> 354234 </td><td>保持无菌</td></tr><tr><td> PBS </td><td> Sigma Aldrich公司</td><td> 100M-8202 </td><td> 10倍的准备凝胶溶液，1个洗涤步骤</td></tr><tr><td>氢氧化钠，1.0N解决方案</td><td> Sigma Aldrich公司</td><td> S2770 </td><td>保持无菌</td></tr><tr><td> DMEM培养液1X </td><td> CellGro </td><td> 10-013-CV </td><td>保持无菌</td></tr><tr><td>胎牛血清</td><td>亚特兰大生物</td><td> 511150 </td><td>保持无菌</td></tr><tr><td>青霉素链霉素</td><td> CellGro </td><td> 30002CI </td><td>保持无菌</td></tr><tr><td>的Triton X-100 </td><td> Sigma Aldrich公司</td><td> X100-500毫升</td><td>在PBS的0.5％ </td></tr><tr><td>多聚甲醛</td><td> Fisher Scientific则</td><td> 04042-500 </td><td> 4％的PBS </td></tr><tr><td>去离子水</td><td></td><td></td><td>保持无菌</td></tr><tr><td> 4&#39;，,6-diaminido-2-苯基吲哚（DAPI） </td><td> MP的Biomedicals </td><td> 0215757401 </td><td> 1毫克/毫升原液</td></tr><tr><td>安装解决方案</td><td> Thermo Scientific的</td><td> TA-030调频</td><td></td></tr><tr><td>胰蛋白酶EDTA </td><td> CellGro </td><td> 25-052-CI </td><td>保持无菌</td></tr></tbody> &lt;/ TABLE&gt;

three-dimensional cell culture model for measuring the effects of interstitial fluid flow on tumor cell invasion

大多数实体肿瘤的生长和发展依赖于初始变换的癌细胞，他们的反应基质的相关信号在肿瘤微环境1。此前，在肿瘤微环境的研究主要集中在肿瘤基质的相互作用1-2。然而，肿瘤微环境，还包括各种生物物理力量，其影响仍然知之甚少。这些部队是肿瘤生长的生物力学的后果，导致基因表达的变化，细胞分裂，分化和入侵3。矩阵密度4，5-6，刚度和结构6-7，组织间液压力8，8间质流体流动都改变在癌症的发展。 在肿瘤间质流体流动特别是高于正常组织8-10。估计间质液FLOW速度进行了测量，发现是在0.1-3微米S -1的范围内，根据肿瘤大小和分化9，11。这是由于肿瘤诱导的血管生成和血管通透性增加12所造成的升高间质流体压力。间质流体流动已被证明能增加入侵的癌细胞13-14血管成纤维细胞和平滑肌细胞15。这次入侵可能是由于自体细胞趋化细胞周围建立3 - D 16的梯度或增加15基质金属蛋白酶（MMP）的表达，分泌趋化因子和细胞粘附分子表达17。然而，细胞感受到流体流动的机制，不能很好地理解。此外，改变肿瘤细胞的行为，间质流体流动的调节其他细胞在肿瘤微环境的活动。它与（一）驾驶的成纤维细胞分化成肿瘤促进myofibr州18，（B）抗原的搬运和其他可溶性因子，淋巴结肿大19，（三）调节淋巴管内皮细胞形态20。 这里介绍的技术对体外细胞间质流体流动和量化入侵（ 图1）的影响。这种方法已经公布在基质和癌细胞侵袭13日至15日，17多个研究测量流体流量的影响。通过改变基质组成，细胞类型和细胞浓度，此方法可以适用于其他疾病和生理系统的研究侵袭，分化，增殖，基因表达，如细胞过程间流动的影响。

Watch this Scientific Journal Video about Three-dimensional Cell Culture Model for Measuring the Effects of Interstitial Fluid Flow on Tumor Cell Invasion at JoVE.com

Three-dimensional Cell Culture Model for Measuring the Effects of Interstitial Fluid Flow on Tumor Cell Invasion

在实体肿瘤间质流体流动升高，可以调节肿瘤细胞的侵袭。在这里，我们描述了技术应用嵌入在一个矩阵的细胞间质流体流动，然后测量其细胞浸润的影响。这种技术可以很容易地适应学习其他系统。

三维细胞培养模式的测量肿瘤细胞浸润间质流体流动的影响

The growth and progression of most solid tumors depend on the initial transformation of the cancer cells and their response to stroma-associated signaling ...

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