Table 2. Advances in sheep ovarian tissue vitrification
Reference Tissue Vitrification device Vitrification solution Main outcomes
Al-aghbari; Menino, 2002. Ovarian fragments (0.5cm x 0.5 cm) Tissue dropped on the surface of a steel cube cooled by LN2 35% EG, 5% polyvinylpyrrolidone, 0.4 mol/L trehalose and 20% FBS Maintenance of oocyte recovery rate and percentage of oocytes developing to metaphase
Bordes et al., 2005. Ovarian fragments (1 mm x 2 cm x 1 cm) Cryogenic vials plunged directly into LN2 2.62 mol/L DMSO, 2.60 mol/L acetamide, 1.31 mol/L 1.2 PROH, and 0.0075 mol/L PEG (gradual dehydration in: 12.5%, 25%, 50% and 100% of vitrification solution) Pregnancies occurred after ovarian cortex autotransplantation, 4 lambs were born
Courbiere et al., 2005. Whole ovaries with vascular pedicle Samples plunged directly into LN2 2.75 mol/L DMSO, 2.76 mol/L formamide, and 1.97 mol/L PROH (gradual dehydration in: 12.5%, 25%, 50% and 100% of vitrification solution) Maintaining primordial follicle viability comparable to the fresh tissue
Courbiere et al., 2006. Whole ovaries Ethyl vinyl acetate cryobag plunged into LN2 2.75 mol/L DMSO, 2.76 mol/L formamide, and 1.97 mol/L PROH (gradual dehydration in: 12.5%, 25%, 50% an 100% of vitrification solution) Maintaining the aspect of the ovarian vein identical to the nonvitrified control
Lornage et al., 2006. Ovarian fragments (1 mm thick and 1 cm2 surface) Cryotube plunged into LN2 2.62 mol/L DMSO, 2.60 mol/L acetamide, 1,31 mol/L 1.2 PROH, and 0.0075 mol/L PEG Pregnancies occurred after autotransplantation of vitrified warmed ovarian cortex and lambs were born
Baudot et al., 2007. Whole ovary with blood vessels. Ethyl vinyl acetate cryobag plunged into LN2 2.7 5 mol/L DMSO, 2.76 mol/L formamide and 1.97 mol/L PROH (gradual dehydration in: 12.5%, 25%, 50% and 100% of vitrification solution) Maintaining ovarian primordial follicle density and follicle membrane integrity similar to the fresh tissue
Courbiere et al., 2009. Whole ovary with blood vessels Ethyl vinyl acetate cryobag plunged into LN2 2.75 mol/L DMSO, 2.76 mol/L formamide, and 1.97 mol/L PROH (gradual dehydration in: 12.5%, 25%, 50% and 100% of vitrification solution) One sheep recovered ovarian endocrine function 6 months after transplantation
Fathi et al., 2011. Ovarian fragments (1 x 2 x 2 mm3) Cryopin (sample adhered in a needle) plunged into LN2 60% HTCM, 15% EG, 15% DMSO, 0.25 mol/L SUC and 10% HSA Maintaining intact antral follicles using 2-step dehydration protocol
Melo et al., 2011. Ovarian fragments (1 mm3) Solid-surface plunged into LN2 40% EG, 0.5 mol/L SUC and 50 μg/mL AA Maintaining follicular viability rates similar to the fresh tissue after 5 days of tissue culture
Lunardi et al., 2012. Ovarian fragments (3 x 3 x 1: 9 mm3) Macrotube plunged into LN2 6 mol/L EG, 0.25 mol/L SUC and 10% FCS Maintaining follicular viability similar to the cultured non- vitrified tissue after 2 days of tissue culture
Torre et al., 2012. Whole ovaries Ethyl vinyl acetate cryobags plunged into LN2 2.75 mol/L DMSO, 2.76 mol/L formamide and 1.97 mol/L PROH (gradual dehydration in: 12.5%, 25%, 50% and 100% of vitrification solution) Significant reduction in ovarian pedicle metabolism after vitrification
Note: LN2 = Liquid nitrogen, EG = Ethylene glycol, FBS = Fetal bovine serum, DMSO = Dimethylsulfoxyde, PROH =: Propylene glycol, PEG = Polyethylene glycol, HTCM = HEPES tissue culture medium, SUC = Sucrose, HSA = Human serum albumin, AA = Ascorbic acid, FCS = Fetal calf serum.