The effectiveness of cryopreservation by vitrification has been reported in many organisms. However, successful silkworm preservation by this method has not been reported. To achieve this, we applied the V-cryoplate method for the cryopreservation of Bombyx mori embryos, a method originally developed in the field of plant cryopreservation. We performed surgery to remove the egg shells and attached the embryos that were enclosed within the serosal membranes to an aluminum cryoplate. Solution exchanges were applied by transferring the cryoplates. Unfortunately, many eggs were crushed and no embryos hatched after cryopreservation. Therefore, we modified the V-cryoplate protocol by mounting paper to the cryoplates with dechorionated eggs, and then removed the mounting paper from the cryoplate before immersion in liquid nitrogen. This modification avoided damage to the serosal membranes and made hatching possible. However, the larvae that hatched from this vitrification cryopreservation died without eating mulberry leaves (the diet of silkworm larvae). These results show that even though embryogenesis mostly progresses normally, some damage occurs. We could improve the method for vitrification for silkworm. However, there are still problems in achieving normal larval development.
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