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

Reconhecemos o apoio financeiro da Ciências Naturais e Pesquisa de Engenharia Council of Canada e da Canadian Institutes of Health Research (CHRPJ 323533-06), o programa de Bolsas de Pós-Graduação Ontário a CM, e os presidentes do Canadá Pesquisa em Micro e Nano Engenharia de Sistemas de YS, e em Mechanobiology a CAS.

<html> A. descrição de dispositivos e operação Dispositivos são fabricados utilizando litografia macia multicamadas 1 em PDMS (polidimetilsiloxano), e são capazes de, simultaneamente, gerando uma série de condições mecânicas em locais de células individuais cultura entre a matriz microfabricated. Neste protocolo, os passos para fabricar um conjunto de pneumático são acionadas microposts descrita pela primeira vez, seguido por etapas para modificar o dispositivo para perm...

<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>

Embora conceitualmente simples, a fabricação de dispositivo faz um pouco de habilidade experimental e prática. Particularmente no caso de cultura de células em 2D, o alinhamento das camadas múltiplas em que o dispositivo pode ser desafiador, especialmente sobre uma matriz de grande área. Praticamente falando, nós podemos confiantemente atingir uma taxa de sucesso de 100% de alinhamento usando dispositivos com 50 mm de tolerância do espaçamento entre as características adjacentes em múltiplas camadas. Temos ta...

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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

A habilidade de sondar sistematicamente in vitro resposta celular a combinações de estímulos mechanobiological para engenharia de tecidos, descoberta de drogas ou estudos biológicos fundamentais célula é limitado pelo biorreator tecnologias atuais, que não pode, simultaneamente, aplicar uma variedade de estímulos mecânicos nas células em cultura. , A fim de resolver este problema, temos desenvolvido uma série de plataformas microfabricated projetado para tela para os efeitos dos estímulos mecânicos em um formato de alto rendimento. Neste protocolo, demonstramos a fabricação de uma matriz microactuator de posts verticalmente deslocadas em que a tecnologia é baseada, e ainda demonstrar como esta tecnologia de base pode ser modificada para conduzir alto rendimento de cultura de células mecanicamente dinâmica em ambos os bidimensionais e três dimensional paradigmas cultura.

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

Microfabricated Platforms for Mechanically Dynamic Cell Culture

Neste protocolo, demonstramos a fabricação de uma matriz microactuator de posts verticalmente deslocadas em que a tecnologia é baseada, e como essa tecnologia base pode ser modificado para conduzir alto rendimento de cultura de células mecanicamente dinâmica em ambos cultura bidimensional e tridimensional paradigmas.

Plataformas microfabricated para cultura de células mecanicamente dinâmico

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

Fabricação de hidrogéis micropatterned para Sistemas de Cultura Neural usando fotolitografia Projeção Dinâmica Máscara

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

Bioengenharia

Shape Memory Polymers for Active Cell Culture

Forma Polímeros Memória da Cultura célula ativa

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

Sistemas Célula de rotação de cultura para cultura de células humanas: células trofoblásticas Humanos como um modelo

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 detecção de fluorescência para as plataformas de gotículas microfluídicos

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

Microscopia correlativo para Análise Estrutural 3D de interações dinâmicas

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

Estudo da migração celular em Canais microfabricados

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

Ambiente Microfluidic controlada de Investigação dinâmico de Agregação Glóbulo Vermelho

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

Fabricação de Mecanicamente Tunable e Bioativos metal andaimes para aplicações 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

Fabricação de espumas de Matriz Extracelular derivados e Microcarregadores como cultura celular de tecidos específicos e 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

Construção de uma folha de células-tronco mesenquimais em várias camadas com um sistema de cultura dinâmica 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 ...