Document Detail


Precise control over the oxygen conditions within the Boyden chamber using a microfabricated insert.
MedLine Citation:
PMID:  20689862     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Cell migration is a hallmark of cancer cell metastasis and is highly correlated with hypoxia in tumors. The Boyden chamber is a porous membrane-based migration platform that has seen a great deal of use for both in vitro migration and invasion assays due to its adaptability to common culture vessels and relative ease of use. The hypoxic chamber is a current tool that can be implemented to investigate the cellular response to oxygen paradigms. Unfortunately, this method lacks the spatial and temporal precision to accurately model a number of physiological phenomena. In this article, we present a newly developed microfabricated polydimethylsiloxane (PDMS) device that easily adapts to the Boyden chamber, and provides more control over the oxygenation conditions exposed to cells. The device equilibrates to 1% oxygen in about 20 min, thus demonstrating the capabilities of a system for researchers to establish both short-term continuous and intermittent hypoxia regimes. A Parylene-C thin-film coating was used to prevent ambient air penetration through the bulk PDMS and was found to yield improved equilibration times and end-point concentrations. MDA-MD-231 cells, an invasive breast cancer line, were used as a model cell type to demonstrate the effect of oxygen concentration on cell migration through the Boyden chamber porous membrane. Continuous hypoxia downregulated migration of cells relative to the normoxic control, as did an intermittent hypoxia regime (IH) cycling between 0% and 21% oxygen (0-21% IH). However, cells exposed to 5-21% IH exhibited increased migration compared to the other conditions, as well as relative to the normoxic control. The results presented here show the device can be utilized for experiments implementing the Boyden chamber for in vitro hypoxic studies, allowing experiments to be conducted faster and with more precision than currently possible.
Authors:
Shawn C Oppegard; Alexander J Blake; Justin C Williams; David T Eddington
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2010-08-05
Journal Detail:
Title:  Lab on a chip     Volume:  10     ISSN:  1473-0197     ISO Abbreviation:  Lab Chip     Publication Date:  2010 Sep 
Date Detail:
Created Date:  2010-08-25     Completed Date:  2010-10-13     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101128948     Medline TA:  Lab Chip     Country:  England    
Other Details:
Languages:  eng     Pagination:  2366-73     Citation Subset:  IM    
Affiliation:
Department of Bioengineering, University of Illinois at Chicago, USA.
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MeSH Terms
Descriptor/Qualifier:
Cell Hypoxia
Cell Line, Tumor
Cell Movement*
Diffusion
Dimethylpolysiloxanes
Humans
Membranes, Artificial*
Microtechnology / methods*
Neoplasm Invasiveness
Oxygen / metabolism*
Polymers / chemistry
Porosity
Xylenes / chemistry
Grant Support
ID/Acronym/Agency:
R21 NS051580-01A1/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/Dimethylpolysiloxanes; 0/Membranes, Artificial; 0/Polymers; 0/Xylenes; 25722-33-2/parylene; 63148-62-9/baysilon; 7782-44-7/Oxygen

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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