Biology Reference
In-Depth Information
28
Fluorescence Assays to Study Cell Adhesion
and Migration In Vitro
Paola Spessotto, Emiliana Giacomello, and Roberto Perris
1. Introduction
1.1. Cell Adhesion as the Basis
for the More Complex Process of Cell Migration
The basis for cell movement and the maintenance of organized epithelial
structures formed by “stationary” cells is a phenomenon known as cell adhesion,
i.e., the establishment of the firm anchorage of a cell to an underlying substra-
tum. Both cell adhesion and the subsequent process of cell locomotion occur
via the interaction of cell-adhesion molecules, integrins, cell-surface
proteoglycans, nonintegrin receptors, and selections with each other or with
extracellular ligands such as various ECM constituents. The number of extra-
cellular matrix (ECM) components discovered to be implicated in cell adhe-
sion and migration phenomena is rapidly growing. This fact, in conjunction
with the progressively unveiled diversity in the mode by which these ECM
molecules affect cell behavior, has raised the demands on the in vitro assays
aimed at the analysis of cell adhesion. Probably, the requirement for a higher
accuracy and versatility; for an increased capability to most closely reproduce
the in vivo situations; and for the uncompromised possibility to apply the assays
to the analysis of ex vivo cells; represent the primary demands that have forced
investigators to ameliorate the currently available cell-adhesion protocols, as
well as device novel alternative ones.
In order to move, a cell needs to adhere to the underlying substratum by a
reversible and adequately strong binding
(1
,
2)
. Thus, in some instances the
adhesive interactions could be strong, whereas in others it could be relatively
weak, as for instance in the case of rapidly locomoting cells and circulating
lymphocytes. Accordingly, for a comprehensive understanding of the mecha-
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