The use of eryopreservation for convenient bioassay of insect organs in vitro

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Abstract
Pheromone glands of the female moth Helicoverpa (Heliothis) armigera and Malpighian tubules of the locust Locusta migratoria migratorioides (R&F) retained partial biological activity after eryopreservation. Pheromone glands were frozen to ‐70oC in a tissue culture medium (TC‐199) containing 10% of the cryoprotectant dimethylsulfoxide. Malpighian tubules, however, required initial preincubation for 24 h in a medium of high osmolarity before being frozen to ‐70oC in the presence of 10% of the cryoprotectant glycerol. After rapid thawing of both organs, biological activity was compared with the respective fresh organs. Using a radiochemical assay, it was shown that previously frozen pheromone glands were stimulated by pheromone biosynthesis activating neuropeptide to the same degree as fresh pheromone glands. In addition, previously frozen Malpighian tubules were stimulated by an extract of corpora cardiaca to produce c‐AMP to the same extent as fresh Malpighian tubules.