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

Wij erkennen de financiële steun van het Natural Sciences and Engineering Research Council van Canada en de Canadese Institutes of Health Research (CHRPJ 323533-06), de Ontario Graduate Scholarship programma CM, en de Canada Onderzoek Stoelen in Micro en Nano Engineering Systems aan YS, en in Mechanobiology naar CAS.

<html> A. Device beschrijving en werking Apparaten worden vervaardigd met behulp van meerlaagse soft lithografie 1 in polydimethylsiloxaan (PDMS), en zijn in staat van gelijktijdige opwekking van een scala van mechanische omstandigheden in individuele cel cultuur locaties in het hele microfabricated array. In dit protocol zijn de stappen om een ​​reeks van pneumatisch aangedreven-micropalen fabriceren voor het eerst beschreven, gevolgd door stappen om het apparaat te wijzigen om ...

<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>

Hoewel conceptueel eenvoudig, het apparaat fabricage vergt wel wat experimentele vaardigheid en oefening. Vooral in het geval van een 2D-celkweek, kan uitlijning van de meerdere lagen in het apparaat een uitdaging zijn, vooral over een grote oppervlakte array. Praktisch gezien kunnen we betrouwbaar bereiken van een 100% afstemming succes gebruik van apparaten met 50 urn van tolerantie in de ruimte tussen aangrenzende functies in meerdere lagen. We hebben ook met succes aangetoond afstemming met toleranties zo laag als 1...

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<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

De mogelijkheid om systematisch onderzoek in vitro cellulaire respons op combinaties van mechanobiological stimuli voor tissue engineering, drug discovery of fundamentele celbiologie studies wordt beperkt door de huidige bioreactor technologieën, die niet tegelijkertijd van toepassing zijn een verscheidenheid aan mechanische stimuli aan gekweekte cellen. Om dit probleem aan te pakken, hebben we een serie van microfabricated platforms ontwikkeld om de effecten van mechanische stimuli scherm in een high-throughput formaat. In dit protocol, tonen we de fabricage van een Microactuator reeks van verticaal verschoven berichten op de technologie gebaseerd is, en verder aan te tonen hoe deze basistechnologie kunnen worden gewijzigd om high-throughput mechanisch dynamische celkweek te voeren in zowel twee-dimensionale en drie- dimensionale cultuur paradigma&#39;s.

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

Microfabricated Platforms for Mechanically Dynamic Cell Culture

In dit protocol, tonen we de fabricage van een Microactuator reeks van verticaal verschoven berichten op de technologie gebaseerd is, en hoe deze basistechnologie kunnen worden gewijzigd om high-throughput mechanisch dynamische celkweek te voeren in zowel twee-dimensionale en drie-dimensionale cultuur paradigma&#39;s.

Microfabricated Platforms voor Mechanisch Dynamic Cel Cultuur

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

Fabricage van micropatterned Hydrogelen voor Neurale Cultuur Systems met behulp van Dynamic Mask Projectie Fotolithografie

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

Shape Memory Polymers for Active Cell Culture

Shape Memory Polymeren voor actieve cel Cultuur

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

Roterende celkweek systemen for Human Cultuur van de Cel: Human trofoblast cellen als een model

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

Fluorescentie detectiemethoden voor microfluïdische druppel platforms

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

Correlatieve microscopie voor 3D Structural Analysis van Dynamic Interactions

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

Studie van cel-migratie in Microfabricated Kanalen

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

Gecontroleerde Microfluïdische Milieu voor Dynamic Onderzoek van Red Blood Cell Aggregation

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

Fabricage van mechanisch afstembare en bioactieve Metal Steigers voor biomedische toepassingen

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

Fabricage van extracellulaire matrix-afgeleide schuimen en microdragers Weefsel-specifieke Cell Cultuur en Delivery Platforms

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

Bouw van een meerdere lagen mesenchymale stamcellen blad met een 3D dynamische cultuur-systeem

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 ...