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

Noi riconosciamo il sostegno finanziario della scienze naturali e ingegneria Research Council del Canada e del Canadian Institutes of Health Research (CHRPJ 323533-06), la borsa di studio Ontario Graduate programma a CM, ei presidenti canadese Research in Micro e Nano Sistemi Ingegneria di YS, e in Mechanobiology al CAS.

<html> A. Descrizione del dispositivo e il funzionamento I dispositivi sono fabbricati con multistrato litografia soft 1 in polidimetilsilossano (PDMS), e sono in grado di generare simultaneamente una serie di condizioni meccaniche in singoli luoghi di coltura cellulare attraverso l&#39;array microfabbricazione. In questo protocollo, i passaggi per fabbricare una vasta gamma di azionamento pneumatico-microposts sono descritta per la prima, seguita dalla procedura per modificare il di...

<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>

Anche se concettualmente semplice, fabbricazione del dispositivo ci vuole una certa abilità sperimentale e pratica. In particolare nel caso di colture cellulari in 2D, l&#39;allineamento degli strati multipli nel dispositivo può essere impegnativo, soprattutto in una grande area array. In pratica, possiamo ottenere un affidabile al 100% tasso di successo allineamento utilizzando dispositivi con 50 micron di tolleranza nella spaziatura tra le funzioni adiacenti in strati multipli. Abbiamo anche dimostrato con successo ...

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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 capacità di sondare sistematicamente in risposta cellulare in vitro a combinazioni di stimoli mechanobiological per l&#39;ingegneria tissutale, la scoperta di farmaci o fondamentali studi di biologia cellulare è limitata dalle tecnologie attuali bioreattore, che non può allo stesso tempo applicare una varietà di stimoli meccanici di cellule in coltura. Per affrontare questo problema, abbiamo sviluppato una serie di piattaforme di microfabbricazione progettato per schermo per gli effetti degli stimoli meccanici in un formato high-throughput. In questo protocollo, dimostriamo la realizzazione di una serie di messaggi microattuatore verticalmente sfollati su cui si basa la tecnologia, e illustra, inoltre, come questa tecnologia di base può essere modificata per condurre high-throughput di coltura cellulare dinamica meccanicamente in entrambe le due dimensioni e tre paradigmi cultura dimensionale.

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

Microfabricated Platforms for Mechanically Dynamic Cell Culture

In questo protocollo, dimostriamo la realizzazione di una serie di messaggi microattuatore verticalmente sfollati su cui si basa la tecnologia, e come questa tecnologia di base può essere modificato per condurre high-throughput di coltura cellulare meccanicamente dinamica sia la cultura bidimensionale e tridimensionale paradigmi.

Piattaforme microfabbricazione per coltura cellulare meccanicamente dinamica

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

Fabbricazione di idrogel Micropatterned per i sistemi Cultura neurali utilizzando Dynamic Fotolitografia proiezione Mask

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

Ricerca

Bioingegneria

Shape Memory Polymers for Active Cell Culture

Shape Memory Polymers per la Cultura cella attiva

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

Rotazione Cultura sistemi cellulari per colture cellulari umane: cellule del trofoblasto umano come modello

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

Metodi di rilevamento a fluorescenza per piattaforme goccia microfluidica

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 correlativa per l&#39;analisi strutturale 3D di Interazioni Dinamiche

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

Studio della Migrazione cellulare in canali microfabbricati

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 controllato Microfluidic per Dynamic Indagine Globulo rosso Aggregazione

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

Fabbricazione di meccanicamente Tunable e Bioactive metallo Ponteggi per applicazioni biomediche

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

Fabbricazione di Schiume matrice extracellulare-derivati ​​e microcarriers come tessuto-specifica delle cellule, Cultura e piattaforme di distribuzione

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

Costruzione di un foglio di multistrato delle cellule staminali mesenchimali con un sistema di cultura dinamica 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 ...