Name: a device for performing cell migration/wound healing in a 96-well plate
Uploaded: 2017-03-07T15:00:00
Description: Watch this Scientific Journal Video about A Device for Performing Cell Migration/Wound Healing in a 96-Well Plate at JoVE.com

笔者想感谢谭宝梁先生，王梓健先生和香港浸会大学理学院车间的技术人员，对他们的技术技能和建议，使这一wounder的雏形。

1.简介Wounder的部件（图1） <ol><li>通过在相等的距离固定伤人引脚准备针座。握住枪头，并上下移动调节伤人引脚调节尖端的高度。 </li><li>适合于不同品牌的96孔培养板中的可调节的引导杆，并确保该受伤区域是在一个固定的位置上。 </li><li>通过拧紧六角螺丝固定可调伤人引脚。通过拧紧两端的六角帽固定在适当位置的指导吧。 </li></ol> 2.设置销座的宽度（图2） <ol><li>松开六...

<ol>
	<li>Friedl, P., Wolf, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumour-cell+invasion+and+migration:+diversity+and+escape+mechanisms.">Tumour-cell invasion and migration: diversity and escape mechanisms.</a> Nat Rev Cancer. 3 (5), 362-374 (2003).</li><li>Friedl, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prespecification+and+plasticity:+shifting+mechanisms+of+cell+migration.">Prespecification and plasticity: shifting mechanisms of cell migration.</a> Curr Opin Cell Biol. 16 (1), 14-23 (2004).</li><li>Lampugnani, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+migration+into+a+wounded+area+in+vitro.">Cell migration into a wounded area in vitro.</a> Methods Mol Biol. 96, 177-182 (1999).</li><li>Sholley, M. M., Gimbrone, M. A., Cotran, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+migration+and+replication+in+endothelial+regeneration:+a+study+using+irradiated+endothelial+cultures.">Cellular migration and replication in endothelial regeneration: a study using irradiated endothelial cultures.</a> Lab Invest. 36 (1), 18-25 (1977).</li><li>Gotlieb, A. I., Spector, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Migration+into+an+in+vitro+experimental+wound:+a+comparison+of+porcine+aortic+endothelial+and+smooth+muscle+cells+and+the+effect+of+culture+irradiation.">Migration into an in vitro experimental wound: a comparison of porcine aortic endothelial and smooth muscle cells and the effect of culture irradiation.</a> Am J Pathol. 103 (2), 271-282 (1981).</li><li>Chen, Y. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single-cell+migration+chip+for+chemotaxis-based+microfluidic+selection+of+heterogeneous+cell+populations.">Single-cell migration chip for chemotaxis-based microfluidic selection of heterogeneous cell populations.</a> Sci Rep. 18 (5), 9980 (2015).</li><li>Yarrow, J. C., Periman, Z. E., Westwood, N. J., Mitchison, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+high-throughput+cell+migration+assay+using+scratch+wound+healing,+a+comparsion+of+image-based+readout+methods.">A high-throughput cell migration assay using scratch wound healing, a comparsion of image-based readout methods.</a> BMC Biotechnol. 4, 21 (2004).</li><li>Lauder, H., Frost, E. E., Hiley, C. R., Fan, T. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+the+repair+process+involved+in+the+repair+of+a+cell+monolayer+using+an+in+vitro+model+of+mechanical+injury.">Quantification of the repair process involved in the repair of a cell monolayer using an in vitro model of mechanical injury.</a> Angiogenesis. 2 (1), 67-80 (1998).</li><li>Yue, P. Y. K., Leung, E. P. Y., Mak, N. K., Wong, R. N. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simplified+method+for+quantifying+cell+migration/+wound+healing+in+96-well+plates.">A simplified method for quantifying cell migration/ wound healing in 96-well plates.</a> J. Biomol Screen. 15 (4), 427-433 (2010).</li><li>Yue, P. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Elucidation+of+the+mechanisms+underlying+the+angiogenic+effects+of+ginsenoside+Rg(1)+in+vivo+and+in+vitro.">Elucidation of the mechanisms underlying the angiogenic effects of ginsenoside Rg(1) in vivo and in vitro.</a> Angiogenesis. 8 (3), 205-216 (2005).</li><li>Kwok, H. H., Chan, L. S., Poon, P. Y., Yue, P. Y., Wong, R. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ginsenoside-Rg1+induces+angiogenesis+by+the+inverse+regulation+of+MET+tyrosine+kinase+receptor+expression+through+miR-23a.">Ginsenoside-Rg1 induces angiogenesis by the inverse regulation of MET tyrosine kinase receptor expression through miR-23a.</a> Toxicol Appl Pharmacol. 287 (3), 276-283 (2015).</li></ol>

我们的细胞wounder在解决传统的细胞迁移实验的问题，几个独特的功能。 8通道机械细胞wounder与可以通过热压灭菌寿命长采用优质不锈钢（304）的。几乎所有的商业品牌的市场上可用的96孔培养板可与该机械细胞wounder使用，因为可调节导向杆的。可调导向杆的设计也保证了划痕区域是在每个的中心孔。在井的中心位置上的伤口可以使用显微镜促进图像捕获。图像具有一些变型中，而在不同的时间点...

细胞迁移在细胞过程，如胚胎发生，神经发生，血管生成，伤口愈合，粘膜修复，在正常发育和疾病状况1上皮-间质跃迁中起重要作用。它是一个复杂的过程，涉及大量的间和细胞内的事件，包括信号分子相互作用，细胞极化，细胞骨架重组，基质重塑，膜突起和动态细胞-细胞粘连调制2的协调。通过研究细胞迁移，化学物质或生物分子导致细胞的运动和它的相关的生物化学途径的发现和验证可以被确定。这一基本过程可以有利地用作用于各种应用，如疾病的治疗和药物靶向的代理。 的伤人测定是细胞迁移测定法3中的一个。这里，一个细胞的个体的样品将通过机械装置被部分地去除，以产生一个裸露区，让细胞将迁移以覆盖面积。回收的在特定时间点的百分比将被监测。当处理大量的样品，如样品中实验成本和交叉污染的问题可能会出现问题。虽然许多商业工具和测定法可用于研究细胞迁移4，5，6，其中许多是需要昂贵和复杂的设备和仍需要改进。由于这些原因，一个8通道机械细胞wounder（ 图1）显影。 我们的8通道机械细胞wounder在解决上述问题已经将几个独特的功能。它提供了执行细胞迁移/在96孔培养板格式伤人测定的灵活性。可调区iding杆设计保证了刮开区是在每个中心位置很好。另外，导向杆的高度可以调整，使得wounder适用于各种品牌培养板。最后，可调伤人针设计能够与培养板表面实现同步和可重复伤人7,8的连枪头接触。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>96-well cell culture plate</td>
			<td>Nunc</td>
			<td>167008</td>
			<td>Other brands of 96-well cell culture plate can also be used</td>
		</tr>
		<tr>
			<td>P10 pipette tips</td>
			<td>Axygen</td>
			<td>301-03-051</td>
			<td>Short P10 pipette tip is more easy to create a clear wound</td>
		</tr>
		<tr>
			<td>Wounder</td>
			<td>R&#38;P Technology Limited</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Medium 199</td>
			<td>Sigma</td>
			<td>M2520-1L</td>
			<td>For cell culture of human umbilical vein endothelial cells. 
			Use appropirate culture medium and condition for other type of cells.</td>
		</tr>
		<tr>
			<td>Fetal bovine serum</td>
			<td>Gibco</td>
			<td>26140079</td>
			<td>For cell culture of human umbilical vein endothelial cells. 
			Use appropirate culture medium and condition for other type of cells.</td>
		</tr>
		<tr>
			<td>Penicillin/Streptomycin</td>
			<td>Gibco</td>
			<td>15140122</td>
			<td>For cell culture of human umbilical vein endothelial cells. 
			Use appropirate culture medium and condition for other type of cells.</td>
		</tr>
		<tr>
			<td>Heparin sodium salt from porcine intestinal mucosa</td>
			<td>Sigma</td>
			<td>H3393</td>
			<td>For cell culture of human umbilical vein endothelial cells. 
			Use appropirate culture medium and condition for other type of cells.</td>
		</tr>
		<tr>
			<td>Gelatin from bovine skin</td>
			<td>Sigma</td>
			<td>G9391</td>
			<td>For cell culture of human umbilical vein endothelial cells. 
			Use appropirate culture medium and condition for other type of cells.</td>
		</tr>
	</tbody>
</table>

a device for performing cell migration/wound healing in a 96-well plate

细胞迁移/伤人测定法是一种常用的方法来研究细胞迁移和其它生物过程，如血管生成和肿瘤转移。在该试验中，细胞生长，以形成汇合单层，并通过与一个设备刮擦创建的机械伤口。然后朝裸露区域的细胞的迁移速率可以通过成像来监测。我们的8通道机械wounder旨在解决大部分与细胞迁移测定相关的问题。首先，我们的wounder可以容易地通过高压灭菌或与常用消毒剂消毒。其次，个人可调节销允许甚至与细胞培养板，使得可以创建锐利和可再现的伤口接触。第三，关于wounder两侧的导杆确保在每一致伤人位置很好。为伤人还可以提供更好的处理wounder的以及以最小化交叉铅污染的使用一次性塑料吸头离子。总之，我们的细胞wounder可以使用标准的96孔培养板进行细胞迁移测定法提供与用户友好和可重现装置的研究人员。

该8通道机械wounder被构造以执行小区迁移测定划伤的细胞单层。这是一个用户友好的设备，可以在不到一分钟进行96孔板细胞划伤实验无需特殊培训。此wounder可以在细胞单层引入伤口面积为约600微米的均匀的宽度，并与锋利的边缘（ 图5和6）。伤口的宽度取决于品牌枪头的和用户的性能。伤口宽度的范围适用于普通图像的低功率放大下捕获（ <stron...

Watch this Scientific Journal Video about A Device for Performing Cell Migration/Wound Healing in a 96-Well Plate at JoVE.com

A Device for Performing Cell Migration/Wound Healing in a 96-Well Plate

在这里，我们提出了一个协议，以在96孔细胞培养板进行半高通量细胞迁移测定。该协议是建立在一个细胞单层一致划伤伤口快速，简便，经济的方法。

一种在96孔板表演细胞迁移/伤口愈合设备

The cell migration/wounding assay is a commonly used method to study cell migration and other biological processes, such as angiogenesis and tumor ...

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