Cryoprotectants synergy improve zebrafish sperm cryopreservation and offspring skeletogenesis.

摘要: Abstract The synergy obtained by the combination of cryoprotectants is a successful strategy that can be beneficial on optimization zebrafish sperm cryopreservation. Recently, protocol was established for this species using an electric ultrafreezer (−150 °C) performing cooling rate (−66 °C/min) and storage within one step. ultimate objective cryopreservation to generate healthy offspring. Therefore, study select most adequate cryoprotectant combination, previously protocol, high quality offspring with normal skeletogenesis. Among permeating concentrations studied 12.5% 15% N,N-dimethylformamide (DMF) yielded post-thaw hatching rates. For these two concentrations, presence bovine serum albumin (10 mg/mL), egg yolk (10%), glycine (30 mM) bicine (50 mM) evaluated motility, viability, in vitro fertilization success skeletal development (30 days post fertilization). Higher concentration (15%) decreased incidence deformed arches severe malformations, which suggests higher capacity protect cell against cold stress DNA damage. Extender containing DMF Ctrl, Bicine were non-permeating quality. use compounds results reduction vertebral fusions, compressions severity malformations extender compositions are sired cryopreserved −66 °C/min freezing rate. To best our knowledge, first report performed different media zebrafish.