EVALUATION OF TWO in vitro MATURATION MEDIUM FOR EMBRYO PRODUCTION IN SHEEP

Abstract

The aim of this work was to evaluate the effect of HECM-9 and TCM-199 as maturation media on maturation (MR), in vitro fertilization (IVF) and embryo development (ED) rate of oocytes from hair sheep collected from slaughter house ovaries. Cumulus-oocyte-complexes (COC) were obtained by manual aspiration, from 2-6mm diameter follicles. Groups of 10-20 COC, quality 1 and 2 were placed into 450μL of HECM-9 or TCM-199 and incubated 24h at 38.5 ºC and 5% CO2. For IVF, COC were transferred to 450μL of SOFm+Oaa, 0.5 x 106 motile spermatozoa were added and then incubated at 38.5 ºC and 5% CO2. Alleged zygotes were transferred to 450μL of SOFm+Oaa+glucose. Embryo development and morphology were evaluated at 2, 4, 5 and 6 days of culture, not developed zygotes were removed on day 2 and the final rate of embryo production was determined on day 8 of culture. Oocyte MR showed no significant differences (P>0.05) between treatments (73.3 vs. 71.4 % HECM-9 and TCM-199, respectively). Fertilization rate was different (P<0.05) between treatments, observing a better response from oocytes matured in HECM-9 than those matured in TCM-199 (47.4 vs. 28.6%, respectively). Embryo development was greater (P<0.05) in oocytes matured in HECM-9 than those matured in TCM-199 (26.3 vs. 7.9%, respectively). We conclude that HECM-9 increased in vitro fertilization rate and embryo development in ovine oocytes.