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Preservation of bovine preantral follicle viability and ultra-structure after cooling and freezing of ovarian tissue.
Celestino, Juliana Jales de Hollanda; dos Santos, Regiane Rodrigues; Lopes, Cláudio Afonso Pinho; Martins, Fabrício Sousa; Matos, Maria Helena Tavares; Melo, Mônica Aline Parente; Báo, Sônia Nair; Rodrigues, Ana Paula Ribeiro; Silva, José Roberto Viana; de Figueiredo, José Ricardo.
Afiliação
  • Celestino JJ; Laboratory of Manipulation of Oocytes and Ovarian Preantral Follicles-LAMOFOPA, Faculty of Veterinary, Ceará State University, Fortaleza, CE, Brazil.
Anim Reprod Sci ; 108(3-4): 309-18, 2008 Nov.
Article em En | MEDLINE | ID: mdl-17945440
Bovine preantral follicles within ovarian fragments were exposed and cryopreserved in absence or presence of 1.5M glycerol (GLY), ethylene glycol (EG), propanediol (PROH) or dimethyl sulfoxide (DMSO), undergoing a previous cooling at 20 degrees C for 1h (protocol 1) or at 4 degrees C for 24h (protocol 2) in 0.9% saline solution. At the end of each treatment, preantral follicles were classified as non-viable/viable when they were stained/not stained with trypan blue, respectively. To confirm viability staining, ultra-structure of the follicles was evaluated by transmission electronic microscopy (TEM). Data were compared by Chi-square test (P<0.05). The storage of the ovaries at 20 degrees C for 1h (78%) and 4 degrees C for 24h (80%) did not reduce significantly the percentage of viable preantral follicles when compared to the control (75%). Similar results were obtained when ovarian fragments, respectively, for protocols 1 and 2, were exposed to MEM (78 and 77%), 1.5M EG (78 and 71%), as well as frozen in 1.5M EG (74 and 77%). Percentages of viable follicles in control were similar to those observed after exposure (75%) and freezing (76%) in presence of 1.5M DMSO only when protocol 1 was used. The increase of the concentration from 1.5 to 3.0M, for all cryoprotectants, reduced significantly the percentage of viable preantral follicles after freezing. Ultra-structural analysis has confirmed trypan blue results, showing that not only basement membrane, but also organelles, were intact in viable preantral follicles. In conclusion, ovarian tissue cooling at 4 degrees C for 24h before cryopreservation (protocol 2) does not affect the viability of bovine preantral follicles when 1.5M EG is present in the cryopreservation medium.
Assuntos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Bovinos / Criopreservação / Crioprotetores / Folículo Ovariano Tipo de estudo: Guideline Limite: Animals Idioma: En Revista: Anim Reprod Sci Ano de publicação: 2008 Tipo de documento: Article País de afiliação: Brasil País de publicação: Holanda
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Bovinos / Criopreservação / Crioprotetores / Folículo Ovariano Tipo de estudo: Guideline Limite: Animals Idioma: En Revista: Anim Reprod Sci Ano de publicação: 2008 Tipo de documento: Article País de afiliação: Brasil País de publicação: Holanda