Abstract

FERNANDEZ REYES, F; HERNANDEZ PICHARDO, J.E  and  PICHARDO CRUZ, A.I. In vitro MATURATION OF BOVINE OOCYTES WITH FSH+LH AND FSH IN CULTURE MEDIUM. Rev Salud Anim. [online]. 2007, vol.29, n.2, pp. 105-110. ISSN 0253-570X.

For in vitro maturation of ovine oocytes, ovaries from slaughtered hybrid animals were obtained. Immediately after slaughter, ovaries were placed on physiological saline solution at 35^oC. A total of 242 ovaries was collected and 1294 follicles with a size between 2 to 6 mm in diameter were obtained. From them, 107 oocytes showing 3 or more compact layers of cumulus cells and homogenous cytoplasm were selected and cultivated in a maturation medium TCM 199 supplemented with FSH and LH in concentrations of: 1) 5mg/mL of FSH and 5mg/mL of LH (22 oocytes). 2) 2.5 mg/mL of each FSH and LH (16 oocytes). 3) 5mg/mL of FSH (36 oocyres), 4) 2.5 mg/mL of FSH (33 oocytes). Subsequently, they were fixed and evaluated by the acetic orceine staining. The maturation results indicate that the group supplemented with FSH 2.5 mg/mL, showed the highest percentage (45.4%) of Metaphase II oocytes, followed by the group supplemented with 5 mg/mL of FSH (27.7%) and the group supplemented with 2.5 mg/mL of FSH + 2.5mg/mL LH (22.7%). Significant statistic differences among the groups were not observed (P< 0.05%). According to these results, it is concluded that the best medium for the ovine oocytes in vitro maturation is TCM-99 supplemented with FSH 2.5 mg/mL. The in vitro manipulation of ovine oocytes may allow the establishment of reproductive programs in the natural crossing age period and also out of it. Genetic material banks for several applications may also be established.

Keywords : in vitro maturation; oocytes; ovine; FSH; L.H.

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