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The re-suspended cells are then transferred to a 1.8-ml Eppendorf tube containing the same freezing medium and a 30-50% layer of glycerol at -20 degrees C. Prior to the freezing step, the cells are incubated for 2 h in a medium free of DMSO. The cells are cryopreserved by slow freezing in 10% dimethyl sulfoxide (DMSO) at -80 degrees C. The authors present here a protocol for cryopreservation of these cells, as well as for long-term storage of trout pituitary secretions.
Trout gonadotrophs are widely used for the study of the neuroendocrine regulation of sexual development, reproductive physiology and the conservation of this important vertebrate model.
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