Effects of different sucrose concentrations on vitrified porcine preantral follicles: Qualitative and quantitative analysis

GABRIEL, P.; TORRES, P.; FRATTO, C.; CISALE, H.; CLAVER, J.A.; LOMBARDO, D.M.; FISCHMAN, ML.

CRYOBIOLOGY

ACADEMIC PRESS INC ELSEVIER SCIENCE

Lugar: Amsterdam; Año: 2017 vol. 76 p. 1 - 7

0011-2240

The aim of the present study was to perform a qualitative and quantitative analysis of the effect ofdifferent sucrose concentrations combined with ethylene glycol in the preservation of vitrified porcinepreantral follicles. Fragments of ovarian cortex were vitrified in cryotubes containing 200 ml of thevitrification solution (30% Ethylene Glycol; 20% Fetal Bovine Serum; 0 Me0.25 M e 0.75 M or 1 M sucrose)and stored in liquid nitrogen for a week. Histological analysis showed that after vitrification thenumber of normal follicles decreased compared to the fresh tissue (control). The percentage of normalprimordial follicles was sucrose dose dependent. The percentage of normal primary follicles was similarin 0 M or 0.25 M sucrose, while higher concentrations (0.75 M and 1 M) increased significantly thepercentage of abnormal follicles (p < 0.05). Morphometric analysis showed a statistically significantreduction in the total area of primordial follicles with 0.75 M sucrose and a significant increase in thecytoplasmic area of primordial follicles with 0 M sucrose (p < 0.05). The qualitative and the quantitativeanalysis appear to be a complementary tool when choosing a vitrification protocol. For our cryopreservationsystem - vitrification of ovarian cortex slices in cryotubes-the best vitrification medium wasTCM 199-Hepes with 30% de ethylene glycol, 20% of Fetal Bovine Serum and 0 or 0.25 M sucrose. Thepresent study shows that the use of high sucrose concentrations in the vitrification solution has adeleterious effect on the preservation of porcine preantral follicles contained in ovarian tissue. Consequently,its use at 0.75 M or 1 M wouldn´t be recommended.