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

We acknowledge financial support from the Natural Sciences and Engineering Research Council of Canada and the Canadian Institutes of Health Research (CHRPJ 323533-06), the Ontario Graduate Scholarship program to CM, and the Canada Research Chairs in Micro and Nano Engineering Systems to YS, and in Mechanobiology to CAS.

A. Device description and operation
Devices are fabricated using multilayer soft lithography1 in polydimethylsiloxane (PDMS), and are capable of simultaneously generating a range of mechanical conditions in individual cell culture locations across the microfabricated array. In this protocol, the steps to fabricate an array of pneumatically-actuated microposts are first described, followed by steps to modify the device to enable mechanically dynamic culture in both two-dimensional (...

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Though conceptually simple, device fabrication does take some experimental skill and practice. Particularly in the case of 2D cell culture, alignment of the multiple layers in the device can be challenging, especially over a large-area array. Practically speaking, we can reliably achieve a 100% alignment success rate using devices with 50 &mu;m of tolerance in spacing between adjacent features in multiple layers. We have also successfully demonstrated alignment with tolerances as low as 15 &mu;m, but the alignment tim...

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		<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

The ability to systematically probe in vitro cellular response to combinations of mechanobiological stimuli for tissue engineering, drug discovery or fundamental cell biology studies is limited by current bioreactor technologies, which cannot simultaneously apply a variety of mechanical stimuli to cultured cells. In order to address this issue, we have developed a series of microfabricated platforms designed to screen for the effects of mechanical stimuli in a high-throughput format. In this protocol, we demonstrate the fabrication of a microactuator array of vertically displaced posts on which the technology is based, and further demonstrate how this base technology can be modified to conduct high-throughput mechanically dynamic cell culture in both two-dimensional and three-dimensional culture paradigms.

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

Microfabricated Platforms for Mechanically Dynamic Cell Culture

In this protocol, we demonstrate the fabrication of a microactuator array of vertically displaced posts on which the technology is based, and how this base technology can be modified to conduct high-throughput mechanically dynamic cell culture in both two-dimensional and three-dimensional culture paradigms.

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

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

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Fluorescence detection methods for microfluidic droplet 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

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

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

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

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

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

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 ...