351 EFFECTS OF FETAL CALF SERUM, GROWTH FACTOR, AND HORMONE SUPPLEMENTATION DURING IN VITRO MATURATION ON PARTHENOGENETIC ACTIVATION AND EMBRYO DEVELOPMENT OF FOLLICULAR RABBIT OOCYTES
Z. Polgar A , T. Somfai A B , V. Angeli A , X. H. Tang C , W. Ji C and A. Dinnyes A DA Genetic Reprogramming Group, Agricultural Biotechnology Center, Godollo, Hungary
B Genetic Diversity Department, National Institute of Agrobiological Sciences, Tsukuba, Japan
C Kunming Institute of Zoology, CAS, Kunming, Yunnan, P.R. of China
D Research Group for Applied Animal Genetics and Biotechnology, HAS and Szent Istvan University, Godollo, Hungary
Reproduction, Fertility and Development 19(1) 290-291 https://doi.org/10.1071/RDv19n1Ab351
Submitted: 12 October 2006 Accepted: 12 October 2006 Published: 12 December 2006
Abstract
Improvement of the in vitro maturation (IVM) system for rabbit oocytes could play a role in rabbit biotechnology. Our goal was to improve IVM to have an efficient source of rabbit oocytes for further studies on nuclear transfer. The effects of FCS, growth factors, and hormone supplementation on oocyte maturation, activation, and embryo development rates were evaluated. Rabbit ovaries were transferred from the slaughterhouse to the laboratory in PBS. Oocytes were collected by aspiration and subjected to IVM in 3 types of media based on TCM-199. In one group, growth factors (50 ng mL−1 of insulin-like growth factor-I and 10 ng mL−1 of epidermal growth factor) and hormones (5 IU mL−1 of hCG and 5 IU mL−1 of pregnant mare serum gonadotropin) and BSA were added to the IVM medium (IVM+); in the other groups, IVM medium was supplemented with either 10% (IVM + 10% FCS) or 20% FCS (IVM + 20% FCS). Maturation was assessed by the presence of a polar body after 16 h. Matured oocytes were activated twice by electric stimuli (3 DC pulses, 1.6 kV cm−1, 60 µs) and twice by chemical activation (incubation with 2.5 mM 6-DMAP for 30 min, second time for 2 h) and cultured in vitro in Earle's balanced salt solution complete medium at 38.5°C under 5% CO2 in air. Cleavage rates were recorded 16 h after activation and the blastocyst rates were recorded at Day 5 of in vitro culture. Data were analyzed by ANOVA. Maturation rates did not differ between the treatment groups (Table 1). There was no significant difference in cleavage rates between the IVM+ and the IVM + 10% FCS groups; however, the cleavage rate of the IVM + 20% FCS group was significantly lower compared with the others (Table 1; P < 0.05). Development to the blastocyst did not differ significantly between the treatment groups (Table 1; P < 0.05). The results showed that high (20%) FCS supplementation during IVM had a detrimental effect on oocyte cleavage. Hormonal and growth factor supplementation had no beneficial effects on maturation, activation rates, or blastocyst formation, and in fact were not essential for in vitro embryo production in the rabbit.
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The project was supported by RABIOTECH OMFB-00330/2004, EU FP6 (MEXT-CT-2003-509582, and 518240), Wellcome Trust (Grant No. 070246), and TET CH-28/04.
