title: Immobilization of electroporated cells for fabrication of cellular biosensors: physiological effects of the shape of calcium alginate matrices and foetal calf serum.
abstract: In order to investigate the physiological effect of transfected cell immobilization in calcium alginate gels, we immobilized electroporated Vero cells in gels shaped either as spherical beads or as thin membrane layers. In addition, we investigated whether serum addition had a positive effect on cell proliferation and viability in either gel configuration. The gels were stored for four weeks in a medium supplemented or not with 20% (v/v) foetal calf serum. Throughout a culture period of four weeks, cell proliferation and cell viability were assayed by optical microscopy after provision of Trypan Blue. Non-elaborate culture conditions (room temperature, non-CO(2) enriched culture atmosphere) were applied throughout the experimental period in order to evaluate cell viability under less than optimal storage conditions. Immobilization of electroporated cells was associated with an initially reduced cell viability, which was gradually increased. Immobilization was associated with maintenance of cell growth for the duration of the experimental period, whereas electroporated cells essentially died after a week in suspension culture. Considerable proliferation of immobilized cells was observed in spherical alginate beads. In both gel configurations, addition of serum was associated with increased cell proliferation. The results of the present study could contribute to an improvement of the storability of biosensors based on electroporated, genetically or membrane-engineered cells.
authors.affiliation: Laboratory of Plant Physiology, Faculty of Agricultural Biotechnology, Agricultural University of Athens, Greece.
authors.forename: Nikos
authors.initials: N
authors.lastname: Katsanakis
authors.forename: Andreas
authors.initials: A
authors.lastname: Katsivelis
authors.forename: Spiridon
authors.initials: S
authors.lastname: Kintzios