Reduced polyspermic fertilization of porcine oocytes utilizing elevated bicarbonate and reduced calcium concentrations in a single-medium system
J. R. Herrick A , M. L. Conover-Sparman A and R. L. Krisher A B
+ Author Affiliations
- Author Affiliations
A Department of Animal Sciences, 915 W State St, Lilly Hall of Life Sciences, Purdue University, West Lafayette, IN 47907-1151, USA.
B Author to whom correspondence should be addressed. email: rkrisher@purdue.edu
Reproduction, Fertility and Development 15(4) 249-254 https://doi.org/10.1071/RD03001
Submitted: 2 January 2003 Accepted: 23 May 2003 Published: 23 May 2003
Abstract
The development of efficient systems for in vitro production of porcine embryos has been hampered by a high incidence of polyspermic fertilization. A recently developed single-medium system for porcine in vitro maturation (IVM), IVF and in vitro embryo culture (IVC) (Purdue Porcine Medium; PPM) was modified with elevated bicarbonate (44 mM) and reduced calcium concentrations (1.7 mM) for IVF (PPMfert.2). Oocyte penetration was evaluated after maturation in PPMmat (0.5 mg mL−1 hyaluronan, 0.6 mM cysteine, 10 ng mL−1 epidermal growth factor (EGF), 0.1 U mL−1 porcine LH and FSH, and 1 × Minimal Essential Medium (MEM) vitamins) and fertilization (5 h with 5 × 105 sperm mL−1) in either PPMfert.2 or mTBM (20 mM Tris, 0.0 mm bicarbonate, 7.5 mM calcium). Embryonic development (cleavage and blastocyst stages) was assessed after culture in PPM1 and PPM2. Although penetration was lower in PPMfert.2 (69.9%) compared with mTBM (83.9%), 48.8% of penetrated oocytes were fertilized normally in PPMfert.2 compared with only 27.8% normal fertilization in mTBM. More oocytes cleaved in PPMfert.2 (77.9% v. 53.7%), but development to the blastocyst stage was not different between treatments (14.1% v. 14.3%). Further work is needed to improve embryonic development, but reduced polyspermic penetration is an important step in the optimization of the PPM system for in vitro porcine embryo production.
Extra keywords: capacitation
Acknowledgments
We thank Indiana Packers for their kind donation of ovarian tissue and A. Brad, K. Stroble, A. Clifford and R. Durkin for the transportation of ovaries to the laboratory. Dr W. Singleton is also thanked for his comments on an early version of this manuscript.
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