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1.
Biophys J ; 98(9): 1790-800, 2010 May 19.
Article in English | MEDLINE | ID: mdl-20441742

ABSTRACT

We report quantitative measurements of the velocity field of collectively migrating cells in a motile epithelium. The migration is triggered by presenting free surface to an initially confluent monolayer by using a microstencil technique that does not damage the cells. To avoid the technical difficulties inherent in the tracking of single cells, the field is mapped using the technique of particle image velocimetry. The main relevant parameters, such as the velocity module, the order parameter, and the velocity correlation function, are then extracted from this cartography. These quantities are dynamically measured on two types of cells (collectively migrating Madin-Darby canine kidney (MDCK) cells and fibroblastlike normal rat kidney (NRK) cells), first as they approach confluence, and then when the geometrical constraints are released. In particular, for MDCK cells filling up the patterns, we observe a sharp decrease in the average velocity after the point of confluence, whereas the densification of the monolayer is much more regular. After the peeling off of the stencil, a velocity correlation length of approximately 200 microm is measured for MDCK cells versus only approximately 40 microm for the more independent NRK cells. Our conclusions are supported by parallel single-cell tracking experiments. By using the biorthogonal decomposition of the velocity field, we conclude that the velocity field of MDCK cells is very coherent in contrast with the NRK cells. The displacements in the fingers arising from the border of MDCK epithelia are very oriented along their main direction. They influence the velocity field in the epithelium over a distance of approximately 200 microm.


Subject(s)
Cell Movement , Epithelial Cells/cytology , Animals , Cell Line , Dogs , Epithelial Cells/metabolism , Models, Biological , Molecular Imaging , Rats , Time Factors
2.
Proc Natl Acad Sci U S A ; 104(41): 15988-93, 2007 Oct 09.
Article in English | MEDLINE | ID: mdl-17905871

ABSTRACT

Using an original microfabrication-based technique, we experimentally study situations in which a virgin surface is presented to a confluent epithelium with no damage made to the cells. Although inspired by wound-healing experiments, the situation is markedly different from classical scratch wounding because it focuses on the influence of the free surface and uncouples it from the other possible contributions such as cell damage and/or permeabilization. Dealing with Madin-Darby canine kidney cells on various surfaces, we found that a sudden release of the available surface is sufficient to trigger collective motility. This migration is independent of the proliferation of the cells that mainly takes place on the fraction of the surface initially covered. We find that this motility is characterized by a duality between collective and individual behaviors. On the one hand, the velocity fields within the monolayer are very long range and involve many cells in a coordinated way. On the other hand, we have identified very active "leader cells" that precede a small cohort and destabilize the border by a fingering instability. The sides of the fingers reveal a pluricellular actin "belt" that may be at the origin of a mechanical signaling between the leader and the followers. Experiments performed with autocrine cells constitutively expressing hepatocyte growth factor (HGF) or in the presence of exogenous HGF show a higher average velocity of the border and no leader.


Subject(s)
Cell Movement/physiology , Wound Healing/physiology , Animals , Cell Communication/drug effects , Cell Communication/physiology , Cell Culture Techniques , Cell Line , Cell Movement/drug effects , Cell Polarity , Cell Shape , Dogs , Epithelial Cells/cytology , Epithelial Cells/physiology , Hepatocyte Growth Factor/pharmacology , Hepatocyte Growth Factor/physiology , Microscopy, Fluorescence , Models, Biological , Signal Transduction/physiology
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