Effect of different support systems in the preservation of vitrified porcine preantral follicles contained in ovarian cortex slices: preliminary results

GABRIEL P; CISALE H; FRATTO C; LOMBARDO DM; CLAVER JA; FISCHMAN ML

Lyon

Congreso; 18º International Congress on Animal Reproduction; 2016

IETS

Effect of different support systems in the preservation of vitrified porcine preantral follicles contained in ovarian cortex slices: preliminary results.Gabriel, P.1; Fratto, M.C.1; Claver, J.A.2; Cisale H.1; Lombardo D2.; Fischman M.L.1 Cátedras de 1Física Biológica y 2Histología y Embriología. INITRA. Facultad de Cs. Veterinarias. Universidad de Buenos Aires. Argentina. fischman@fvet.uba.arOvarian tissue vitrification allows the preservation of a large quantity of oocytes contained in preantral follicles (PAF). In previous work, we found that ethylene glycol (E) was the less harmful cryoprotective agent (CPA) [1]. The aim of the present study was to analyze the effect of different support systems in the preservation of porcine PAF histologic structure during the vitrification process using E and sucrose as CPA. Slaughter porcine ovaries were used (n=5). Eight samples of the ovarian cortex (5mm x 2mm x 1mm) were taken from each ovary. Controls were fixed in Bouin solution. The remaining samples were exposed to a vitrification solution (VS: TCM-199, 25Mm HEPES and antibiotic; 30% E; 20% Fetal Bovine Serum and 0.25M sucrose). Two samples were vitrified in cryotubes containing 200 μL of VS (Treatment A). Two were placed in BEEM® capsules (Treatment B) and the remaining in acupuncture needles (Treatment C), both systems using minimum volume of VS. Samples were stored in liquid nitrogen for one week. Histological evaluation was performed by haematoxylin-eosin staining and observed at light microscope (x400). PAF where classified in primordial and primary according to the standards established by Smitz [2] and then classified as normal (without alterations in the oocyte or in the granulosa cells) or abnormal (with degenerative changes such as injured nuclear membrane, shrinkage of nucleus, cytoplasm vacuolization or eosinophilia, granulosa cells with high pycnosis or disruption of the epithelium) [3]. A total of 285 PAF were analyzed. Statistical analysis was performed using the non-parametric Friedman test (P