Isolation and culture of pig tonsil lymphocytes

Tonsils are secondary lymphoid organs that play an important role in host defense. The aim of our study was to develop reliable procedures for isolation and culture of pig tonsil cells, and to validate their possible use in functional immunoassays. Using our isolation procedure, we recovered on average 238.7 ± 107.1 × 106 cells per tonsil couple with a mean vitality of 89.8 ± 2.7%. These values significantly decreased 8 months after freezing at −80 °C along with the subsequent spontaneous release of both IgA and IgG in culture. These results suggest to use pig tonsil cells within 2 months from thawing to maintain suitable conditions in terms of recovery, vitality and release of antibody in vitro. Tonsil mononuclear cells also showed the ability to secrete antimicrobial peptides and to respond in vitro to immunological stimuli. On the whole, our study has defined operating conditions for tonsil processing, control of bacterial contaminations, time limits of storage at −80 °C, as well as for evaluating polyclonal Ig production in vitro. Such procedures are likely to be of some importance in studies on regional immunity and in the development of large animal models for biomedical sciences.