Abstract
The ability of macrophages to migrate to sites of infection and inflammation is critical for their role in the innate immune response. Macrophage cell lines have made it possible to study the roles of individual proteins responsible for migration using molecular biology, but it has not been possible to reliably elicit the motility of macrophage cell lines in two dimensions. In the past, measurements of the motility of macrophage cell lines have been largely limited to transwell assays which provide limited quantitative information on motility and limited ability to visualize cell morphology. We used microcontact printing to create polydimethylsiloxane (PDMS) surfaces functionalized with fibronectin that otherwise support little macrophage adhesion. We used these surfaces to measure macrophage migration in two dimensions and found that these cells migrate efficiently in a uniform field of colony-stimulating factor-1, CSF-1. Knockdown of Cdc42 led to a nonstatistically significant reduction in motility, whereas chemical inhibition of PI3K activity led to a complete loss of motility. Inhibition of the RhoA kinase, ROCK, did not abolish the motility of these cells but caused a quantitative change in motility, reducing motility significantly on high concentrations of fibronectin but not on low concentrations. This study illustrates the importance of studying cell motility on well controlled materials to better understand the exact roles of specific proteins on cell migration.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 542-554 |
| Number of pages | 13 |
| Journal | Cytoskeleton |
| Volume | 71 |
| Issue number | 9 |
| DOIs | |
| State | Published - Sep 1 2014 |
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Keywords
- Fibronectin
- Macrophage
- Microcontact printing
- Motility
- Podosome
ASJC Scopus subject areas
- Cell Biology
- Structural Biology
Cite this
Two-dimensional motility of a macrophage cell line on microcontact-printed fibronectin. / Hind, Laurel E.; Mackay, Joanna L.; Cox, Dianne; Hammer, Daniel A.
In: Cytoskeleton, Vol. 71, No. 9, 01.09.2014, p. 542-554.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Two-dimensional motility of a macrophage cell line on microcontact-printed fibronectin
AU - Hind, Laurel E.
AU - Mackay, Joanna L.
AU - Cox, Dianne
AU - Hammer, Daniel A.
PY - 2014/9/1
Y1 - 2014/9/1
N2 - The ability of macrophages to migrate to sites of infection and inflammation is critical for their role in the innate immune response. Macrophage cell lines have made it possible to study the roles of individual proteins responsible for migration using molecular biology, but it has not been possible to reliably elicit the motility of macrophage cell lines in two dimensions. In the past, measurements of the motility of macrophage cell lines have been largely limited to transwell assays which provide limited quantitative information on motility and limited ability to visualize cell morphology. We used microcontact printing to create polydimethylsiloxane (PDMS) surfaces functionalized with fibronectin that otherwise support little macrophage adhesion. We used these surfaces to measure macrophage migration in two dimensions and found that these cells migrate efficiently in a uniform field of colony-stimulating factor-1, CSF-1. Knockdown of Cdc42 led to a nonstatistically significant reduction in motility, whereas chemical inhibition of PI3K activity led to a complete loss of motility. Inhibition of the RhoA kinase, ROCK, did not abolish the motility of these cells but caused a quantitative change in motility, reducing motility significantly on high concentrations of fibronectin but not on low concentrations. This study illustrates the importance of studying cell motility on well controlled materials to better understand the exact roles of specific proteins on cell migration.
AB - The ability of macrophages to migrate to sites of infection and inflammation is critical for their role in the innate immune response. Macrophage cell lines have made it possible to study the roles of individual proteins responsible for migration using molecular biology, but it has not been possible to reliably elicit the motility of macrophage cell lines in two dimensions. In the past, measurements of the motility of macrophage cell lines have been largely limited to transwell assays which provide limited quantitative information on motility and limited ability to visualize cell morphology. We used microcontact printing to create polydimethylsiloxane (PDMS) surfaces functionalized with fibronectin that otherwise support little macrophage adhesion. We used these surfaces to measure macrophage migration in two dimensions and found that these cells migrate efficiently in a uniform field of colony-stimulating factor-1, CSF-1. Knockdown of Cdc42 led to a nonstatistically significant reduction in motility, whereas chemical inhibition of PI3K activity led to a complete loss of motility. Inhibition of the RhoA kinase, ROCK, did not abolish the motility of these cells but caused a quantitative change in motility, reducing motility significantly on high concentrations of fibronectin but not on low concentrations. This study illustrates the importance of studying cell motility on well controlled materials to better understand the exact roles of specific proteins on cell migration.
KW - Fibronectin
KW - Macrophage
KW - Microcontact printing
KW - Motility
KW - Podosome
UR - http://www.scopus.com/inward/record.url?scp=84907877365&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84907877365&partnerID=8YFLogxK
U2 - 10.1002/cm.21191
DO - 10.1002/cm.21191
M3 - Article
C2 - 25186818
AN - SCOPUS:84907877365
VL - 71
SP - 542
EP - 554
JO - Cytoskeleton
JF - Cytoskeleton
SN - 1949-3584
IS - 9
ER -