In vitro development of bovine embryos cultured with stem cell factor or insulin-like growth factor-I following IVF with semen of two bulls having different field fertility
Introduction
The success of AI depends on many factors. The estimated relative conception rate (ERCR) is a measure of the fertility of an individual sire (Clay and McDaniel, 2001). The ERCR predicted fertility value is a 70-d non-return rate of an AI service relative to the service sires of herd mates. Many laboratory procedures have been used to test different aspects of semen quality to predict field fertility. Yet, the usefulness of these techniques remains inconclusive (Rodriguez-Martinez, 2003). The ability of sperm to fertilize oocytes following in vitro fertilization (IVF) differs among bulls (Lonergan, 1994, Shi et al., 1990) and sire used in IVF has a profound effect on in vitro embryonic development (Ward et al., 2003).
The in vitro culture environment has a great influence on the development and quality of embryos. Though in vitro embryo production using serum supplemented media is widely accepted, serum contains many undefined factors that may stimulate or inhibit embryo development (Yamashita et al., 1999). Development may vary with serum from different sources (Batt and Miller, 1988), and serum has a biphasic effect on embryo development in cattle. Serum can inhibit the first cleavage division, but it can stimulate blastocyst development (Van Langendonckt et al., 1997). Experiments have been attempted to investigate the effects of growth factors on embryonic development in cattle. Growth factors improved embryo development, but they have not yielded results comparable to fetal bovine serum (FBS). However, previous research reported that the use of growth factors such as epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I) improved the rate of blastocyst formation of embryos. Kurzawa et al. (2004) demonstrated that EGF and IGF-I, at a concentration of 100 ng/mL, had a positive effect on mouse embryos. Culture medium supplemented with IGF-I was beneficial for preimplantation embryo development because it increased cell proliferation in human, rabbit and mice blastocysts (Makarevich and Markkula, 2002). With embryo culture in cattle, EGF supplementation was less effective that IGF-I or stem cell factor (SCF, Dhali et al., 2007b). Also, it is recognized that oocyte quality and the development competence following IVF is determined by the characteristics of follicles and follicular size (Qian et al., 2001).
The overall hypotheses were that growth factor supplementation to culture was equal to an undefined medium containing FBS for embryo development and in the absence of serum supplement field fertility differences could be evaluated with in vitro embryo development. The objective was to investigate the in vitro development of embryos in cattle after IVF with the semen of bulls of above and below average field fertility. For the precise evaluation of the sire effects on embryo development, a total serum-free system was used for in vitro embryo production to exclude the possible undesirable influence of serum on embryo development.
Section snippets
Chemicals
All the chemicals used in the experiments were procured from Mallinckrodt Baker Inc., Phillipsburg, NJ, USA unless mentioned otherwise. The IGF-I, SCF, essential amino acids solution, non-essential amino acids solution, tissue culture medium 199 (TCM 199) and gentamicin sulfate solution were procured from Invitrogen Corporation, Carlsbad, CA, USA. The FBS, BSA (Fraction V; A9647), FSH, LH, 17β-estradiol, sodium pyruvate, l-glutamine, heparin and phenol red solution were procured from
Results
The variations in cleavage rate, blastocyst formation, expanded/hatched blastocyst formation and embryo development score in different culture media by bull and follicle size are given in Table 2. The overall embryo development for the two main effects of bull and follicle size is depicted in Fig. 1. Cleavage rate did not differ significantly between the bulls. Simultaneously the different IVC conditions did not have significant influences on cleavage rate. Nevertheless, the effect of follicle
Discussion
The experiment was conducted to assess the in vitro development of embryos in cattle derived with the semen of bulls of above and below average field fertility. Oocyte maturation and IVF were performed in serum-free media and IVC was in serum supplemented or growth factor supplemented serum-free medium. The results indicate that the in vitro embryo development did not differ between the bulls, though their field fertility performance was different.
In vitro embryo development may be influenced
Acknowledgment
The authors wish to thank Select Sire Power Inc. for donation of semen for these studies.
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