In Vitro Maturation, InVitro Fertilization, and Embryonic Development of Canine Oocytes

Abstract

We investigated the efficiency of invitro mauturation (IVM) as a basic experiment to study the development of canine oocytes after invitro fertilization (IVF). Therefore, we performed two-part experiments. First, experiment I compared the effects of TCM 199 without fetal bovine serum (FBS) to TCM 199 supplemented with 5 % FBS on the invitro nuclear maturation rate of canine oocytes. To investigate the efficiency of meiotic development to the metaphase II (MII) stages, 4.7 % (4/64) of all oocytes in TCM 199 without FBS developed to the MII stage compared with 1.7% (1/59) in TCM 199 with 5 % FBS for 48h. FBS did not increase invitro nuclear maturation. In experiment II, the cleavage rate of canine oocytes was investigated after heparin treatment on IVF. Canine oocytes were fertilized as four groups: Fert-TALP medium without heparin (FertI) or Fert-TALP medium supplemented with10,20,30ug/ml heparin (Fert II, Fert III, Fert IV, respectively). Oocytes for 24 h fertilization in Fert I were the highest of all of the groups as 6.5% (5/77) in Fert I, and developed to early morula. Especially, the oocytes cultured in Fert I for 24 h of insemination time had a higher rate of embryonic development than other groups. Therefore, we can assert that unlike in other mammals, heparin in canine IVF does not increase the efficiency of fertilization rate, and is not an important factor.