
pmid: 11034794
Prior studies on cryopreserving embryos of several non-drosophilid flies established that two Drosophila melanogaster embryo cryopreservation protocols were not directly suitable for use with these species. This paper describes our work on developing a protocol for cryopreservation of embryos of the housefly, Musca domestica. Significant progress was made when permeabilization of the vitelline membrane was optimized, a vitrification solution containing ethylene glycol, polyethylene glycol, and trehalose was formulated, and when cooling and recovery of the cryopreservation protocol included a step which passed the embryos through liquid nitrogen vapor. More than 70% of housefly embryos withstand treatments of dechorionation, permeabilization, loading with cryoprotectant, and dehydration in vitrification solution, but the cooling, warming, and poststorage rearing steps still cause a considerable reduction in survival. About 53% of the vitrified M. domestica embryos hatched into larvae. Relative to the percentage of the control adult emergence, about 13% of the embryos stored in liquid nitrogen developed into fertile adults. Hatching of the F(1) progeny of adults having been cryopreserved as embryos was similar to control levels.
Cryopreservation, Male, Ethylene Glycol, Embryo, Nonmammalian, Nitrogen, Lipids, Permeability, Culture Media, Heptanes, Polyethylene Glycols, 2-Propanol, Solutions, Cryoprotective Agents, Fertility, Houseflies, Animals, Hexanes, Female, Desiccation, Chromatography, High Pressure Liquid
Cryopreservation, Male, Ethylene Glycol, Embryo, Nonmammalian, Nitrogen, Lipids, Permeability, Culture Media, Heptanes, Polyethylene Glycols, 2-Propanol, Solutions, Cryoprotective Agents, Fertility, Houseflies, Animals, Hexanes, Female, Desiccation, Chromatography, High Pressure Liquid
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