Name: microfabricated platforms for mechanically dynamic cell culture
Uploaded: 2010-12-26T17:00:00
Description: Watch this Scientific Journal Video about Microfabricated Platforms for Mechanically Dynamic Cell Culture at JoVE.com

Reconocemos el apoyo financiero de las Ciencias Naturales e Ingeniería de Investigación de Canadá y los Institutos Canadienses de Investigación en Salud (CHRPJ 323533-06), el programa de Becas de Posgrado Ontario a CM, y los presidentes de Investigación en Micro y Nano Sistemas de Ingeniería de YS, y en Mecanobiología al CAS.

<html> A. Descripción de dispositivo y la operación Dispositivos se fabrican utilizando la litografía blanda multicapa en un polidimetilsiloxano (PDMS), y son capaces de generar al mismo tiempo una serie de condiciones mecánicas en los distintos lugares de cultivo de células a través de la matriz microfabricated. En este protocolo, los pasos para fabricar una amplia gama de neumático accionado por microposts se describió por primera vez, seguido de pasos para modificar el dis...

<ol>
	<li>Unger, M. A., Chou, H. P., Thorsen, T., Scherer, A., Quake, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monolithic+microfabricated+valves+and+pumps+by+multilayer+soft+lithography.">Monolithic microfabricated valves and pumps by multilayer soft lithography.</a> Science. 288, 113-116 (2000).</li><li>Schaffer, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Device+for+the+application+of+a+dynamic+biaxially+uniform+and+isotropic+strain+to+a+flexible+cell+culture+membrane.">Device for the application of a dynamic biaxially uniform and isotropic strain to a flexible cell culture membrane.</a> J Orthop Res. 12, 709-719 (1994).</li><li>Jo, B., Lerberghe, L. V. a. n., Motsegood, K., Beebe, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+micro-channel+fabrication+in+polydimethylsiloxane+(PDMS)+elastomer.">Three-dimensional micro-channel fabrication in polydimethylsiloxane (PDMS) elastomer.</a> Journal of Microelectromechanical Systems. 9, 76-81 (2000).</li><li>Moraes, C., Sun, Y., Simmons, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Solving+the+shrinkage-induced+PDMS+alignment+registration+issue+in+multilayer+soft+lithography.">Solving the shrinkage-induced PDMS alignment registration issue in multilayer soft lithography.</a> J. Micromech. Microeng. 19, 065015-065015 (2009).</li><li>Haubert, K., Drier, T., Beebe, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PDMS+bonding+by+means+of+a+portable,+low-cost+corona+system.">PDMS bonding by means of a portable, low-cost corona system.</a> Lab on a chip. 6, 1548-1549 (2006).</li><li>Bongaerts, J., Fourtouni, K., Stokes, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Soft-tribology:+Lubrication+in+compliant+PDMS-PDMS+contact.">Soft-tribology: Lubrication in compliant PDMS-PDMS contact.</a> Tribology International. 40, 1531-1542 (2007).</li><li>Moraes, C., Chen, J. H., Sun, Y., Simmons, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfabricated+arrays+for+high-throughput+screening+of+cellular+response+to+cyclic+substrate+deformation.">Microfabricated arrays for high-throughput screening of cellular response to cyclic substrate deformation.</a> Lab on a chip. 10, 227-234 (2010).</li><li>Liu, V. A., Bhatia, S. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+photopatterning+of+hydrogels+containing+living+cells.">Three-dimensional photopatterning of hydrogels containing living cells.</a> Biomedical Microdevices. 4, 257-266 (2002).</li><li>Hahn, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photolithographic+patterning+of+polyethylene+glycol+hydrogels.">Photolithographic patterning of polyethylene glycol hydrogels.</a> Biomaterials. 27, 2519-2524 (2006).</li><li>Moraes, C., Wang, G., Sun, Y., Simmons, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+microfabricated+platform+for+high-throughput+unconfined+compression+of+micropatterned+biomaterial+arrays.">A microfabricated platform for high-throughput unconfined compression of micropatterned biomaterial arrays.</a> Biomaterials. 31, 577-584 (2010).</li><li>Wipff, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+covalent+attachment+of+adhesion+molecules+to+silicone+membranes+for+cell+stretching+applications.">The covalent attachment of adhesion molecules to silicone membranes for cell stretching applications.</a> Biomaterials. 30, 1781-1789 (2009).</li><li>Moraes, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrating+polyurethane+culture+substrates+into+poly(dimethylsiloxane)+microdevices.">Integrating polyurethane culture substrates into poly(dimethylsiloxane) microdevices.</a> Biomaterials. 30, 5241-5250 (2009).</li><li>Ifkovits, J. L., Burdick, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review:+photopolymerizable+and+degradable+biomaterials+for+tissue+engineering+applications.">Review: photopolymerizable and degradable biomaterials for tissue engineering applications.</a> Tissue engineering. 13, 2369-2385 (2007).</li></ol>

Aunque conceptualmente simple, la fabricación del dispositivo tiene una cierta habilidad y la práctica experimental. Particularmente en el caso del cultivo de células en 2D, la alineación de las múltiples capas en el dispositivo puede ser un reto, sobre todo en un conjunto de gran superficie. En términos prácticos, de manera fiable puede lograr una tasa de éxito del 100% de alineación uso de dispositivos con 50 micras de tolerancia en el espaciado entre elementos adyacentes en múltiples capas. También se ha d...

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		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Sylgard 184 PDMS Monomer and Crosslinker Kit</td>
<td>&nbsp;</td>
<td>Dow Corning</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>SU-8 masters</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Silanization agent: (tridecafluoro-1, 1, 2, 2-tetrahydrooctyl)-1-trichlorosilane</td>
<td>&nbsp;</td>
<td>United Chemical Technologies</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Foam pads</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Craft supply stores, 1-2 mm thick</td>
</tr>
<tr>
<td>Overhead inkjet transparencies</td>
<td>&nbsp;</td>
<td>Grand &amp; Toy</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Plexiglass plates</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>C-clamp</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>hardware store</td>
</tr>
<tr>
<td>Micromanipulator system</td>
<td>&nbsp;</td>
<td>Siskiyou</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Custom-made vaccum mount</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Vision system, Navitar 12x zoom</td>
<td>&nbsp;</td>
<td>Navitar</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Connecting tubes</td>
<td>&nbsp;</td>
<td>VWR International</td>
<td>Clay Adam Intramedic PE190</td>
<td>&nbsp;</td>
<tr>
<td>Blunt 18G needles</td>
<td>&nbsp;</td>
<td>Smallparts.com / Amazon Inc.</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Eccentric diaphragm micropump</td>
<td>&nbsp;</td>
<td>Schwarzer Precision</td>
<td>SP 500 EC-LC4.5V DC</td>
<td>&nbsp;</td>
<tr>
<td>Solenoid valves</td>
<td>&nbsp;</td>
<td>Pneumadyne</td>
<td>S10MM-30-12-3</td>
<td>&nbsp;</td>
<tr>
<td>Solenoid manifold</td>
<td>&nbsp;</td>
<td>Pneumadyne</td>
<td>MSV10-1</td>
<td>&nbsp;</td>
<tr>
<td>Function generator</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>3-(trimethoxysilyl) propyl methacrylate</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Polyethylene glycol diacrylate 3.4 kDa</td>
<td>&nbsp;</td>
<td>Laysan Bio</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Polyethylene glycol 8 kDa</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Irgacure 2959</td>
<td>&nbsp;</td>
<td>Ciba Specialty Chemicals</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>1-vinyl-2-pyrolidinone</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Standard cell culture reagents</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

microfabricated platforms for mechanically dynamic cell culture

La capacidad de controlar sistemáticamente en respuesta celular in vitro a combinaciones de estímulos mechanobiological para ingeniería de tejidos, el descubrimiento de fármacos o fundamentales los estudios de biología celular está limitado por las tecnologías de biorreactores actual, que no puede, simultáneamente, aplicar una variedad de estímulos mecánicos a las células cultivadas. Con el fin de abordar esta cuestión, hemos desarrollado una serie de plataformas microfabricated diseñado para detectar los efectos de los estímulos mecánicos en un formato de alto rendimiento. En este protocolo, se demuestra la fabricación de una amplia microactuator de mensajes desplazadas verticalmente en que se basa la tecnología y, además, demostrar cómo esta tecnología de base pueden ser modificados para llevar a cabo de alto rendimiento de cultivo de células mecánica dinámica, tanto en dos dimensiones y tres paradigmas de la cultura dimensional.

Watch this Scientific Journal Video about Microfabricated Platforms for Mechanically Dynamic Cell Culture at JoVE.com

Microfabricated Platforms for Mechanically Dynamic Cell Culture

En este protocolo, se demuestra la fabricación de una amplia microactuator de mensajes desplazadas verticalmente en que se basa la tecnología, y cómo esta tecnología de base pueden ser modificados para llevar a cabo de alto rendimiento de cultivo de células mecánica dinámica, tanto en la cultura de dos dimensiones y tres dimensiones paradigmas.

Plataformas microfabricated para el cultivo celular mecánico dinámico

The ability to systematically probe in vitro cellular response to combinations of mechanobiological stimuli for tissue engineering, drug discovery or ...

Research

JoVE Journal

Bioengineering

Fabrication of Micropatterned Hydrogels for Neural Culture Systems using Dynamic Mask Projection Photolithography

Fabricación de hidrogeles micropatterned para sistemas de cultivo neuronal usando fotolitografía dinámica de proyección Máscara

Increasingly, patterned cell culture environments are becoming a relevant technique to study cellular characteristics, and many researchers believe in the ...

Investigación

Bioingeniería

Shape Memory Polymers for Active Cell Culture

Los polímeros con memoria de forma para el cultivo celular activo

Shape memory polymers (SMPs) are a class of "smart" materials that have the ability to change from a fixed, temporary shape to a pre-determined permanent ...

Rotating Cell Culture Systems for Human Cell Culture: Human Trophoblast Cells as a Model

Rotación de sistemas de cultivos celulares de cultivo de células humanas: células humanas trofoblásticas como un modelo de

The field of human trophoblast research aids in  understanding the complex environment established during placentation. Due to the  nature of these ...

Fluorescence detection methods for microfluidic droplet platforms

Métodos de detección de fluorescencia para las plataformas de las gotas de microfluidos

The development of microfluidic platforms for performing chemistry and biology has in large part been driven by a range of potential benefits that ...

Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions

Microscopía correlativa para el análisis estructural en 3D de Interacciones Dinámicas

Cryo-electron tomography (cryoET) allows 3D visualization of cellular structures at molecular resolution in a close-to-physiological state1. However, ...

Study of Cell Migration in Microfabricated Channels

Estudio de la Migración celular en Canales Microfabricated

The method described here allows the study of cell migration under confinement in one dimension. It is based on the use of microfabricated channels, which ...

Controlled Microfluidic Environment for Dynamic Investigation of Red Blood Cell Aggregation

Medio Ambiente Microfluidic controlada de Investigación Dinámica de agregación de glóbulos rojos

Blood, as a non-Newtonian biofluid, represents the focus of numerous studies in the hemorheology field. Blood constituents include red blood cells, white ...

Fabrication of Mechanically Tunable and Bioactive Metal Scaffolds for Biomedical Applications

Fabricación de Mecánicamente sintonizable y Bioactivos metal andamios para Aplicaciones Biomédicas

Biometal systems have been widely used for biomedical applications, in particular, as load-bearing materials. However, major challenges are high stiffness ...

Fabrication of Extracellular Matrix-derived Foams and Microcarriers as Tissue-specific Cell Culture and Delivery Platforms

La fabricación de espumas y microportadores derivados de la matriz extracelular como específica de tejido de cultivo celular y las plataformas de entrega

Cell function is mediated by interactions with the extracellular matrix (ECM), which has complex tissue-specific composition and architecture. The focus ...

Construction of a Multilayered Mesenchymal Stem Cell Sheet with a 3D Dynamic Culture System

Construcción de una hoja de varias capas células madre mesenquimales con un sistema de cultivo dinámico 3D

Stem cell therapy shows a promising future in regenerating injured organ and tissues, and the cell sheet technique has been developed to improve the low ...