Document Detail


Neutrophil migration assay from a drop of blood.
MedLine Citation:
PMID:  19023468     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Neutrophil directional migration in response to chemical gradients, also known as chemotaxis, is one of the key phenomena in the immune responses against bacterial infection. To better study neutrophils chemotaxis, several in vitro assays have been developed that replicate chemotactic gradients around neutrophils isolated from whole blood. One drawback for most of these assays is the lengthy processing of blood required for neutrophils isolation, which can alter the responsiveness of neutrophils compared to the in vivo conditions. To address this limitation, we have designed a microfluidic chip for chemotaxis studies which can use neutrophils isolated on the chip, directly from whole blood. We have tested three different cell adhesion molecules as substrates for neutrophil isolation (P-selectin, E-selectin and fibronectin) and found average capture efficiencies of 20-40 neutrophils/mm2 at optimized concentrations. Subsequent analysis of neutrophil migration in chemoattractant gradients of N-formyl-methyl-leucyl-phenylalanine (fMLP) or Interleukin-8 (IL-8) shows higher average velocities over E-selectin as compared to the P-selectin. Our microfluidic assay uses just a drop of whole blood (<10 microL) for neutrophil isolation and provides a robust platform to perform chemotaxis assays in the competing environment of different chemokines.
Authors:
Nitin Agrawal; Mehmet Toner; Daniel Irimia
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2008-10-30
Journal Detail:
Title:  Lab on a chip     Volume:  8     ISSN:  1473-0197     ISO Abbreviation:  Lab Chip     Publication Date:  2008 Dec 
Date Detail:
Created Date:  2008-11-21     Completed Date:  2009-01-22     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101128948     Medline TA:  Lab Chip     Country:  England    
Other Details:
Languages:  eng     Pagination:  2054-61     Citation Subset:  IM    
Affiliation:
BioMEMS Resource Center, Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Shriners Hospital for Children, and Harvard Medical School, Boston, MA 02129, USA. dirimia@hms.harvard.edu.
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MeSH Terms
Descriptor/Qualifier:
Blood Chemical Analysis*
Cell Migration Assays*
E-Selectin / analysis,  chemistry
Fibronectins / analysis,  chemistry
Humans
Microchemistry*
Microfluidics / instrumentation,  methods*
Neutrophils / chemistry*
P-Selectin / analysis,  chemistry
Grant Support
ID/Acronym/Agency:
P41 EB002503/EB/NIBIB NIH HHS
Chemical
Reg. No./Substance:
0/E-Selectin; 0/Fibronectins; 0/P-Selectin

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


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