Metabolism: a bottleneck in in vitro toxicological test development. The report and recommendations of ECVAM workshop 54.
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Biomedical subjects
Publications and source records attributed to Brighitta Eletti.
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Cells derived from superficial and deep lymph nodes of transgenic mice in which CD40L expression was deregulated were grown in vitro. After 3 months of interleukin 3 or interleukin 12 stimulation, the cells remained interleukin-independent, showed the same in vitro growth characteristics, but LIL3+ cells were tumorigenic when reinoculated in vivo in nude mice, whereas interleukin-12-treated cells did not induce tumors. Our cell lines could provide a useful model to study the perturbation of the homeostasis allowing us to elucidate the role of cytokines as modulators of differentiation in the lymphoproliferative disorders.
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We review recent progress in the development of models for the freshwater teleost gill based on reconstructed flat epithelia grown on permeable filter supports in primary culture. Methods are available for single-seeded insert (SSI) preparations consisting of pavement cells (PVCs) only from trout and tilapia, and double-seeded insert (DSI) preparations from trout, containing both PVCs (85%) and mitochondria-rich cells (MRCs, 15%), as in the intact gill. While there are some quantitative differences, both SSI and DSI epithelia manifest electrical and passive permeability characteristics typical of intact gills and representative of very tight epithelia. Both preparations withstand apical freshwater exposure, exhibiting large increases in transepithelial resistance (TER), negative transepithelial potential (TEP), and low rates of ion loss, but there is only a small active apical-to-basolateral "influx" of Cl(-) (and not of Na(+)). Responses to various hormonal treatments are described (thyroid hormone T3, prolactin, and cortisol). Cortisol has the most marked effects, stimulating Na(+),K(+)-ATPase activity and promoting active Na(+) and Cl(-) influxes in DSI preparations, and raising TER and reducing passive ion effluxes in both epithelia via reductions in paracellular permeability. Experiments using DSI epithelia lacking Na(+) uptake demonstrate that both NH(3) and NH(4)(+) diffusion occur, but are not large enough to account for normal rates of branchial ammonia excretion, suggesting that Na(+)-linked carrier-mediated processes are important for ammonia excretion in vivo. Future research goals are suggested.