Preservation of biodiversity of rare and endangered fish species by deep freezing methods is relevant for modern science. For the first time defrosted reproductive cells of male Acipenser ruthenus frozen by a method of accoustic-mechanical impact with the use of piezoactuators were used for fertilisation of eggs of sterlet Acipenser ruthenus, which is included in the IUCN Red list The quality of defrosted semen was assessed to determine its fertilisation capacity, which was high, the activity of live active cells up to 79%, and the yield of free embryos up to 62%. The offspring obtained using cryopreserved semen had normal embryonic and postembryonic development, no significant differences from the offspring in the control batch were observed. In terms of reactivity of the central nervous system and receptor complex, the cryopreserved offspring sometimes surpassed the control batch in some parameters such as orienting and background activity, response to stimuli. Larvae after transfer to mixed feeding (10th day) did not significantly differ from the control in the character of the central nervous system reaction, except for the reaction to light stimulus. Thus, the use of frozen sperm using a new method allows us to obtain viable early juveniles adapted to life in natural water bodies for biodiversity conservation.