Serum-free culturing of mammalian cells--adaptation to and cryopreservation in fully defined media.

Long term storage of living cells is a central issue in cell biology and medicine. In addition to the cryoprotectant dimethyl sulphoxide (DMSO), foetal bovine serum (FBS) is often added to the freezing medium for the cryoconservation of serum dependent cell lines. FBS, with its high protein content, protects cells against shear forces and gives the medium a desirable osmotic environment with a physiological viscosity.

Production of antibodies to canine IL-1beta and canine TNF to assess the role of proinflammatory cytokines.

IL-1 and TNF are important proinflammatory cytokines implicated in both antimicrobial host defense and pathogenesis of diseases with an immune-mediated and/or inflammatory component. Respective studies in the dog have been hampered by the unavailability of reagents allowing the specific measurement of canine cytokine proteins and the effect of canine cytokine neutralization by Ab. Starting with recombinant canine (rcan) IL-1beta and rcanTNF, four polyclonal antisera and 22 mAb specific for rcanIL-1beta and rcanTNF were generated.

Monoclonal antibodies: Comparative methods for in vitro production.

The growth of antibody producing hybridoma in the abdominal cavity of the mouse with subsequent harvesting of the ascites fluid is a simple method for the production of monoclonal antibodies. However, due to the massive painful stress of the animal, this method is no longer tolerated by the legislator. In search of alternative methods the scientist has the choice between different techniques: Static tissue culture, spin- or roller-cultures as well as the production in bioreactors (automated cell culture devices).