Extra- and intra-cellular ice formation of red seabream (Pagrus major) embryos at different cooling rates

Technical note: Viability and motility of vitrified/thawed primordial germ cell isolated from common carp (Cyprinus carpio) somite embryos1

“…A number of techniques have been tested for the cryopreservation of fish embryos, including the use of different cryoprotectants, immersion times, and devices for immersing the embryos in liquid nitrogen. However, none have resulted in viable embryos or hatching after vitrification/thawing (Cabrita et al, 2006, Edashige et al, 2006, Li et al, 2009). The visible damage on the surface of the embryo ( Figure 1A) suggests that ice crystals formed during cooling (Higaki et al, 2010a).…”

“…Intracellular ice formation plays a central role in cell injury during cooling (Li et al, 2009). A number of techniques have been tested for the cryopreservation of fish embryos, including the use of different cryoprotectants, immersion times, and devices for immersing the embryos in liquid nitrogen.…”

“…In teleosts, sperm has been successfully cryopreserved (Ohta et al, 2001;Kusuda et al, 2005;Cabrita et al, 2009). However, the development of cryopreservation techniques for teleost embryos has received less attention (Cabrita et al, 2009;Li et al, 2009). …”

Effects of Slow Cooling Methods and Vitrification Methods on Red Seabream (Pagrus major) Embryos

“…However, the cryopreservation of fish embryo has not been successful due to its complex multi-compartmental system, large content of water, high sensitivity to chilling, large amount of egg-yolk and low membrane permeability [3,4]. In recent years, many researches have been carried out on cryopreservation protocols [5] and mechanism of cryoinjuries in fish embryos [6].…”

“…We have carried out some researches on red seabream embryos in cryopreservation protocols [15] and mechanism of cryoinjuries [6]. Potential protocols of slowing cooling and vitrification (sorts of cryoprotectant, programs of cooling and thrawing) have been screened [15].…”