Criopreservação de folículos ovarianos pré-antrais suínos

AUTOR(ES)
DATA DE PUBLICAÇÃO

2009

RESUMO

The present work aimed to test different cryoprotectants on cryopreservation of pig ovarian tissue. Pig ovaries (n=3) were collected at a local slaughterhouse. From each ovary 10 cortex slices were taken, and one was immediately fixed (control) and another placed in in vitro culture (IVC control). The other 8 samples were cryopreserved, in pairs, using 4 different cryoprotectants: DMSO (1.5 M), etilene glycol (EG - 1.5 M) glycerol (GLY - 10%) or propanediol (PROH - 1.5 M) with 0.4% sucrose. Samples were slow cooled and stored in liquid nitrogen for 7 days. After thawing and cryoprotectant removing, one sample from each treatment was immediately fixed and the other was placed in in vitro culture (IVC) for 2h and then fixed. Samples were processed for histology and transmission electron microscopy. The percentages of morphologically normal follicles (MNF) in cryopreserved tissue using DMSO (67.04.9), EG (81.81.4) and PROH (55.99.9) were significantly lower (P<0.05) than the observed in fresh control tissue (97.71.2). When ovarian tissue was cryopreserved with GLY no morphologically normal follicle could be found (0%). After IVC, PROH (40.12.9) presented significantly lower percentage of MNF compared to all other treatments and to the control (85.13.3 DMSO IVC; 84.63.4 EG IVC; 97.51.2 control IVC). In general, the ultrastructure of follicles cryopreserved with DMSO or EG, before and after in vitro culture, was similar. Although their ultrastructure was not very different from control follicles, some alterations could be observed. Signs of advanced degeneration were not observed. In conclusion, the cryopreservation of pig ovarian tissue using DMSO or EG allows a good number of MNF.

ASSUNTO(S)

histologia ultra-estrutura histology ciencias biologicas cultivo in vitro crioprotetores in vitro culture ultrastruture cryoprotectant

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