Vasaturo, Angela (2010) Cell migration in anisotropic media. [Tesi di dottorato] (Unpublished)

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A wide range of basic cellular process depends on cell motility that is fundamental for all eukaryotes. The ability of cells to migrate, adhere, and change shape requires most of the time external signals, although few cells respond primarily to internal cues. One of the most interesting and important response to external stimuli is chemotaxis. Chemotaxis, the directional movement of cells according to a concentration gradient of chemicals, is implicated in physiologically relevant phenomena such as inflammatory response, homeostatic circulation, and development and several disorders including infectious and allergic diseases, wound healing, angiogenesis, atherosclerosis, and tumor metastasis. However, despite the ubiquity and importance of chemotaxis, it remains a difficult process to study in vitro. The work carried out in this thesis present a novel chemotaxis assay in 3-D collagen gels in a direct-viewing chamber. Chemotaxis studies require a way to deliver chemicals to cells in a controlled gradient because cells need to be able to sense an increase in concentration of chemokine to direct their motion. In this chemotaxis assay a chemoattractant concentration gradient in the collagen gel sample seeded with cells is generated by diffusion trough a porous membrane. The diffusion process is monitored by fluorescence microscopy of FITC labelled dextran. Cell motion under the action of the chemoattractant gradient is followed by time-lapse video microscopy. Cell tracking is performed off-line by image analysis and the results are expressed in terms of a chemotactic index and velocity. The assay has been tested by using human neutrophils as a model.

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