Automated quantitative analysis of epithelial cell scatter.
Epithelial cell scatter is a well-known in vitro model for the study of epithelial-mesenchymal transition (EMT). Scatter recapitulates many of the events that occur during EMT, including the dissociation of multicellular structures and increased cell motility.Because it has been implicated in tumor invasion and metastasis,much effort has been made to identify the molecular signals that regulate EMT. To better understand the quantitative contributions of these signals, we have developed metrics that quantitatively describe multiple aspects of cell scatter. One metric (cluster size)quantifies the disruption of intercellular adhesions while a second metric (nearest-neighbor distance) quantifies cell dispersion. We demonstrate that these metrics delineate the effects of individual cues and detect synergies between them. Specifically, we find epidermal growth factor (EGF), cholera toxin (CT) and insulin to synergistically reduce cluster sizes and increase nearest-neighbor distances. To facilitate the rapid measurement of our metrics from live-cell images, we have also developed automated techniques to identify cell nuclei and cell clusters in fluorescence images. Taken together, these studies provide broadly applicable quantitative image analysis techniques and insight into the control of epithelial cell scatter, both of which will contribute to the understanding of EMT and metastasis.
Pope
Melissa D
MD
Division of Engineering and Applied Science, California Institute of Technology, Pasadena, California, USA.
Graham
Nicholas A
NA
Huang
Beijing K
BK
Asthagiri
Anand R
AR
D000818:Animals; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D003584:Cytological Techniques / Q000379:methods*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D006801:Humans
D004815:Epidermal Growth Factor