RESUMO
Ovarian cortical fragments (3 x 3 x 1 mm) were exposed to dimethyl sulfoxide (DMSO) in different concentrations for further analysis of cryoprotectant perfusion by applying high-performance liquid chromatography (HPLC) and conventional cryopreservation. This simple perfusion test can predict the efficiency of the cryopreservation procedure.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Criopreservação/métodos , Crioprotetores/farmacologia , Dimetil Sulfóxido/farmacologia , Folículo Ovariano/efeitos dos fármacos , Ovário/efeitos dos fármacos , Sobrevivência de Tecidos/efeitos dos fármacos , Animais , Crioprotetores/administração & dosagem , Dimetil Sulfóxido/administração & dosagem , Relação Dose-Resposta a Droga , Feminino , Cabras , Modelos Animais , Folículo Ovariano/fisiologia , Ovário/fisiologia , Perfusão/métodos , Sobrevivência de Tecidos/fisiologiaRESUMO
The aim of the present study was to determine the amount of dimethyl sulfoxide (DMSO) present in sheep ovarian tissue after exposure to cryoprotectant at different times (5, 10, 20, or 30 min) and at different concentrations (1.0, 1.5, or 2.0 M). To quantify the levels of DMSO in the ovarian tissue, the high-performance liquid chromatography (HPLC) method was applied. In addition, viability of preantral follicles after toxicity test and cryopreservation of ovarian tissue using the above mentioned concentrations of DMSO and exposure times was evaluated. We have observed that the presence of â¼0.6 mg of DMSO into the ovarian tissue may be deleterious to the sheep preantral follicles. In addition, the application of a short exposure time (5 min at 1.5 or 2.0 M DMSO) or low concentration (1.0 M for 10 min) of DMSO successfully preserves sheep preantral follicles following cryopreservation.