Cryopreservation of Persian sturgeon (Acipenser persicus) embryos by DMSO-based vitrificant solutions

Abstract

Vitrification could provide a promising tool for the cryopreservation of fish embryos. To achieve successful cryopreservation, several parameters should be taken into account in the design of a vitrification protocol. In the present study, some relevant factors were investigated (choice of a proper vitrificant solutions and temperature for thawing) using neurulation-stage embryos. Six DMSO-based vitrificant solutions (V1–V6) were tested using a 6-step incorporation protocol. DMSO-based vitrificant solutions contained DMSO + permeable cryoprotectants + nonpermeable cryoprotectants. Embryos were immersed in vitrificant solutions for 7 minutes and directly plunged into liquid nitrogen. After vitrification (−196 °C for 10 minutes), the thawing was performed in a water bath at 0 or 20 °C and then embryos incubated until hatched. Our results demonstrated that some embryos vitrified in 5 of 6 vitrification solutions survived and hatched out, but none survived after vitrification in V2. The highest survival rate (45.45%) was observed in samples frozen with the best vitrificant solution (V6) and thawing combination (20 °C). These results establish that cryopreservation of Persian sturgeon (Acipenser persicus) embryos by DMSO-based vitrificant solutions is possible.