Assessment of adhesion assays for use with keratinocytes

Cell attachment, as a biological process, is an important aspect with respe ct to graft survival and "take". With the ever-increasing use of cultured e pithelial autografts for coverage and re-epithelialization of wounds, the a ssessment of keratinocyte adhesion in vitro has become a more common requir ement in studies involving extracellular matrix proteins and their receptor molecules, Cell adhesion has been cell-documented in immunological researc h, however keratinocyte adhesion has been investigated by manual counting ( using methylene blue) or other less sensitive colorimetric methods, With th e increase in number of fluorogenic probes available, their use as a sensit ive alternative to radioactive labelling has been promoted in the literatur e. This study was carried out to investigate the possibility of using fluor escent probe 5,6-carboxyfluorescein diacetate succidimyl ester to achieve a more standardized assay in the assessment of keratinocyte adhesion. Adhesi on was assessed on extracellular matrix proteins such as fibronectin, colla gen types I & IV and laminin. We concluded that the fluorescent probe might provide greater sensitivity in measuring adhesion, however it may be cytot oxic to keratinocytes. Pre-labelling of keratinocytes may affect cellular f unctions such as adhesion and even proliferation and consequently the probe must be chosen with care.