The effects of Vitamin A administration on the development of vitrified-warmed mouse blastocyst

The purpose of this study was to evaluate the development of vitrified-warmed mouse blastocysts following a period of Vitamin A administration. Four to six weeks old BALB/c mice were given an intraperitoneal injection of either 0.1 ml paraffin oil alone (control, Con) or paraffin oil containing 250 IU of Vitamin A (experiment, Exp). Ten days later the mice were given second paraffin or paraffin Vitamin A injection and an injection of 10 IU equine chorionic gonadotropin (eCG) followed 48 h later by 10 IU human chorionic gonadotropin (hCG). Blastocysts were collected from both groups and randomly divided into non-vitrified (Con 1, Exp 1) and vitrified (Con 2, Exp 2) subgroups. Embryos in the vitrified group were exposed sequentially to two solutions (10% ethylene glycol, 10% DMSO in holding medium (HM: DMEMF12 + 10% FBS) and 20% ethylene glycol, 20% DMSO in HM) before plunging into liquid nitrogen. After warming at 37 °C, cryoprotectants were diluted serially with 0.25 and 0.15 M sucrose solution in HM. The vitrified-warmed and the fresh embryos of the control and the experiment groups were cultured in DMEMF12 with 10% FBS for 72 h. Although, on the first day of culture, the rate of development to the hatched blastocyst was nearly identical between the two vitrified groups (15.8% versus 13%) but after 48 h, the rate of plated embryos was statistically higher in the vitrified Vitamin A than the vitrified control group (63.1% versus 19.6%, P < 0.001). After 48 h, in the non-vitrified groups, the rate of the plated embryos was also significantly higher in the Vitamin A than the control group (70.5% versus 49.3%, P < 0.01). These data provided evidence that systemic administration of Vitamin A may enhance the potential development of blastocysts in culture and is capable to reduce the adverse effects of vitrification at least during the first 2 days of cultivation.