Animal cloning is a promising technology for biodiversity conservation, and its success depends on the recovery of nucleus donor cells. Specifically for collared peccaries, found sometimes in regions that are difficult to access, the storage at 4-6°C of skin tissues would be an alternative for the conservation of genetic material. Therefore, we aimed to evaluate different storage periods and the presence of a nutrient medium at 4-6°C on the recovery of somatic cells from the skin of collared peccaries. To analyze cell recovery rates, ear explants were distributed in non-refrigerated samples and samples refrigerated for 10, 30, and 50 d in the absence or presence of nutrient medium. All explants were analyzed by histologically and cultured. Only the fragments stored for 50 d without medium showed an increase in the total thickness of skin. Moreover, increased storage period, regardless of the presence of medium, increased the halo number and reduced the metabolic activity. After culture, only the fragments stored without medium for 50 d did not yield any somatic cells. Cells recovered from explants stored for 10 d showed similar characteristics to these recovered from non-refrigerated explants, regardless of the presence of medium, including the day at which explants achieved attachment and the total time to reach subconfluence. In conclusion, viable cells can be recovered from somatic tissues of collared peccaries stored for up to 50 d in the presence of medium, and tissues refrigerated for up to 10 d in the presence of medium yielded more viable cells.
RESUMOO objetivo do presente trabalho foi avaliar a influência do corpo lúteo (CL), glândula endócrina temporária presente nos ovários, sobre a recuperação de oócitos imaturos bovinos derivados de ovários de fêmeas postmortem. Para tanto, as seguintes comparações foram realizadas: fêmeas não prenhes com CL vs. fêmeas não prenhes sem CL; fêmeas prenhes com CL vs. fêmeas não prenhes com CL; e ovário com CL vs. ovário sem CL. Assim, ovários foram obtidos em abatedouro e os oócitos recuperados por aspiração folicular. A qualidade oocitária foi avaliada por morfologia convencional e ensaio de azul cresil brilhante (ACB), sendo quantificados oócitos viáveis e não viáveis para cada técnica. Todos os dados foram analisados pelo teste de Fisher (P<0,05). Assim, avaliando fêmeas ou ovários com ou sem CL, nenhuma diferença foi observada nos parâmetros avaliados. Contudo, um maior percentual de oócitos viáveis por morfologia foi obtido de fêmeas prenhes com CL, quando comparado às fêmeas não prenhes com CL. Em conclusão, pode-se afirmar que parâmetros de recuperação e qualidade oocitária não são afetados pela presença de CL, exceto em doadoras prenhes que apresentaram oócitos de melhor qualidade.
PALAVRAS-CHAVE:azul de cresil brilhante, competência oocitária, morfologia convencional.
INFLUENCE OF THE CORPUS LUTEUM ON THE RECOVERY OF BOVINE IMMATURE OOCYTES DERIVED FROM POST-MORTEM FEMALES ABSTRACTThe aim of the present study was to evaluate the influence of the corpus luteum (CL), temporary endocrine gland present on the ovaries, on the recovery of bovine immature oocytes derived from ovaries of post-mortem females. Thun, the following comparisons were performed: non-pregnant females with CL vs. females not-pregnant without CL; pregnant females with CL vs. not-pregnant females with CL; and ovary with CL vs. ovary without CL. So, ovaries were obtained in slaughterhouse and the oocytes recovered by follicular aspiration. The oocyte quality was evaluated by conventional morphology and brilliant cresyl blue assay (BCB), and viable and non-viable oocytes were quantified for each technique. All data were analyzed by Fisher test (P<0.05). Thus, evaluating females or ovaries with or without CL, no difference was observed in the evaluated parameters. Nevertheless, a higher percentage of viable oocytes by morphology were obtained from pregnant females with CL comparing nonpregnant females with CL. In conclusion, it can be stated that it is possible that recovery parameters and oocyte quality are not affected by the presence of CL, except in pregnant donors who presented oocytes of better quality.
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