The European mink is an endangered Mustelidae species and thus requires effective conservation measures, although little is known about reproduction in this species. In particular, preimplantation development has not been studied and, therefore, embryonic development and the growth of embryos was documented in the present study for European mink using light and fluorescent microscopy. Embryos develop in the oviducts and then migrate into the uterus on Day 6 post coitum (p.c.) at the morula stage. Embryos expanded as blastocysts from Day 7 until implantation on Day 12 p.c. Based on these findings, the use of embryo transfer for a conservation programme for the European mink was evaluated. Embryos were flushed from European mink resource females and transferred into the uterine horns of recipient hybrid females (honoriks and nohoriks). These hybrids were obtained by mating European polecat males with European mink females and vice versa. A total of 40 embryos was transferred and 20 live kits were born. The rates of pre- and postnatal survival were 50% and 70%, respectively. Both male and female offspring were lighter at birth in the embryo transfer group compared with naturally born controls, but there was no difference at 3 months of age.

Lipids are among the most abundant and essential cell components. Specifically, cytoplasmic lipid droplets (LDs) play crucial roles in cellular energy homeostasis. The foci of this review are (1) the composition and roles of lipids during oocyte maturation and early embryonic development, (2) possible causes of cryoinjuries in lipid-rich oocytes/embryos, and (3) ways to overcome these detrimental effects. Recent reports show that LDs in oocytes and embryos are not only energy depots but also are active organelles, possessing many other functions. In addition, analysis of the current literature confirms that lipid phase transition followed by phase separation during cryopreservation is one of the major causes of cryodamage in lipid-rich oocytes and embryos. While LDs and cell membranes are sensitive to chilling and freezing conditions, recent advances in vitrification and delipidation of lipid-rich oocytes and embryos partly mitigate cryodamage. The multidisciplinary approach is critical to reveal mechanisms underlying cryodamage and provides a theoretical basis for optimal cryopreservation of lipid-rich oocytes/embryos.

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Embryo and oocyte cryopreservation is a widely used technology for cryopreservation of genetic resources. One limitation of cryopreservation is the low tolerance to freezing observed for oocytes and embryos rich in lipid droplets. We apply Raman spectroscopy to investigate freezing of lipid droplets inside cumulus-oocyte complexes, mature oocytes, and early embryos of a domestic cat. Raman spectroscopy allows one to characterize the degree of lipid unsaturation, the lipid phase transition from the liquid-like disordered to solid-like ordered state, and the triglyceride polymorphic state. For all cells examined, the average degree of lipid unsaturation is estimated as $1.3 (with 520% deviation) double bonds per acyl chain. The onset of the lipid phase transition occurs in a temperature range from À10 to þ4 C and does not depend on the cell type. Lipid droplets in cumulus-oocyte complexes are found to undergo abrupt lipid crystallization shifted in temperature from the ordering of the lipid conformational state. In the case of mature oocytes and early embryos obtained in vitro, the lipid crystallization is broadened. In the frozen state, lipid droplets inside cumulus-oocyte complexes have a higher content of triglyceride polymorphic b and b 0 phases than estimated for mature oocytes and early embryos. For the first time, to our knowledge, the temperature evolution of the phase state of lipid droplets is examined. Raman spectroscopy is proved to be a promising tool for in situ monitoring of the lipid phase state in a single embryo/oocyte during its freezing.