Single layer centrifugation-selected boar spermatozoa are capable of fertilization in vitro
Good quality spermatozoa are important to achieve fertilization, viable embryos and offspring. Single Layer Centrifugation (SLC) through a colloid (Androcoll-P) selects good quality spermatozoa. However, it has not been established previously whether porcine spermatozoa selected by this method maintain their fertility.
The semen was prepared either by SLC or by standard centrifugation (control) and used for in vitro fertilization (IVF) at oocyte:spermatozoa ratios of 1:50; 1:100 and 1:300 (or 4 x 103, 8 x 103 and 24 x 103 spermatozoa/ml) to evaluate their subsequent ability to generate blastocysts. In addition, sperm motility was assessed by computer assisted sperm motility analysis.
Total and progressive motility were significantly higher in sperm samples prepared by SLC compared to uncentrifuged samples. Sperm binding ability, polyspermy, cleavage and blastocyst rates were affected by the oocyte:sperm ratio, but not by sperm treatment.
The use of SLC does not adversely affect the in vitro fertilizing and embryo-generating ability of the selected spermatozoa compared to their unselected counterparts, but further modifications in the IVF conditions would be needed to improve the monospermy in IVF systems. Since SLC did not appear to have a negative effect on sperm fertilizing ability, and may in fact select for spermatozoa with a greater potential for fertilization, an in vivo trial to determine the usefulness of this sperm preparation technique prior to artificial insemination is warranted.
- Suarez, S (2007) Interactions of spermatozoa with the female reproductive tract: inspiration for assisted reproduction. Reprod Fertil Dev 19: pp. 103-110 CrossRef
- Morrell, J, Rodríguez-Martínez, H (2009) Biomimetic techniques for improving sperm quality in animal breeding: a review. The Open Andrology J 1: pp. 1-9
- Henkel, RR, Schill, WB (2003) Sperm preparation for ART. Reprod Biol Endocrinol 1: pp. 108 CrossRef
- Morrell, J, Rodríguez-Martínez, H (2011) Practical applications of sperm selection techniques as a tool for improving reproductive efficiency. Vet Med International.
- Morrell, J, Wallgren, M (2011) Removal of bacteria from boar ejaculates by single layer centrifugation can reduce the use of antibiotics in semen extenders. Anim Reprod Sci 123: pp. 64-69 CrossRef
- Thys, M, Vandaele, L, Morrell, J, Mestach, J, van Soom, A, Hoogewijs, M, Rodriguez-Martinez, H (2009) In vitro fertilizing capacity of frozen-thawed bull spermatozoa selected by single-layer (glycidoxypropyltrimethoxysilane) silane-coated silica colloidal centrifugation. Reprod Domest Anim 44: pp. 390-394 CrossRef
- Morrell, J, Mari, G, Kútvölgyi, G, Meurling, S, Mislei, B, Iacono, E, Rodriguez-Martinez, H (2011) Pregnancies following artificial insemination with spermatozoa from problem stallion ejaculates processed by single layercentrifugation with Androcoll-E. Reprod Domest Anim 46: pp. 642-645 CrossRef
- Morrell, J, Saravia, F, van Wienen, M, Wallgren, M, Rodriguez-Martinez, H (2009) Selection of boar spermatozoa using centrifugation on a glycidoxypropyltrimethoxysilane-coated silica colloid. J Reprod Dev 55: pp. 547-552 CrossRef
- van Wienen, M, Johannisson, A, Wallgren, M, Parlevliet, J, Morrell, J (2011) Single layer centrifugation with Androcoll-P can be scaled-up to process larger volumes of boar semen. ISRN Vet Sci.
- Kruse, R, Dutta, P, Morrell, J (2011) Colloid centrifugation removes seminal plasma and cholesterol from boar spermatozoa. Reprod Fertil Dev 23: pp. 858-865 CrossRef
- Morrell, J, Wallgren, M (2010) Androcoll™-P-Large selects boar spermatozoa with good membrane integrity from the sperm-rich fraction of the ejaculate. Anim Reprod 7: pp. 16-20
- Caballero, I, Vázquez, JM, Gil, MA, Calvete, JJ, Roca, J, Sanz, L, Parrilla, I, García, EM, Rodríguez-Martínez, H, Martínez, E (2004) Does seminal plasma PSP-I/PSP-II spermadhesin modulate the ability of boar spermatozoa to penetrate homologous oocytes in vitro?. J Androl 25: pp. 1004-1012
- Centurion, F, Vazquez, JM, Calvete, JJ, Roca, J, Sanz, L, Parrilla, I, Garcia, EM, Martinez, EA (2003) Influence of porcine spermadhesins on the susceptibility of boar spermatozoa to high dilution. Biol Reprod 69: pp. 640-646 CrossRef
- García, EM, Vázquez, JM, Calvete, JJ, Sanz, L, Caballero, I, Parrilla, I, Gil, MA, Roca, J, Martínez, EA (2006) Dissecting the protective effect of the seminal plasma spermadhesin PSP-I/PSP-II on boar sperm functionality. J Androl 27: pp. 434-443 CrossRef
- Simonsson, A (1994) Nutrient recommendations for pigs [in Swedish]. Swedish University of Agricultural Sciences, SLU Info. Rapporter, Husdjur, Uppsala, Sweden
- Eriksson, B, Vazquez, J, Martinez, E, Roca, J, Lucas, X, Rodriguez-Martinez, H (2001) Effects of holding time during cooling and type of packaging on plasma membrane integrity, motility and in vitro oocyte penetration ability of frozen-thawed boar spermatozoa. Theriogenology 55: pp. 1593-1605 CrossRef
- Petters, R, Wells, K (1993) Culture of pig embryos. J Reprod Fertil Suppl 48: pp. 61-73
- Yoshioka, K, Suzuki, C, Tanaka, A, Anas, I, Iwamura, S (2002) Birth of piglets derived from porcine zygotes cultured in a chemically defined medium. Biol Reprod 66: pp. 112-119 CrossRef
- Gordon I: Laboratory production of cattle embryos. Second edition. Dublin, Ireland: CABI Publishing; 2003. [Biotechnology in Agriculture Series No. 27]
- Matás, C, Coy, P, Romar, R, Marco, M, Gadea, J, Ruiz, S (2003) Effect of sperm preparation method on in vitro fertilization in pigs. Reproduction 125: pp. 133-141 CrossRef
- Coy, P, Gadea, J, Rath, D, Hunter, R (2009) Differing sperm ability to penetrate the oocyte in vivo and in vitro as revealed using colloidal preparations. Theriogenology 72: pp. 1171-1179 CrossRef
- Hunter, R (1991) Oviduct function in pigs, with particular reference to the pathological condition of polyspermy. Mol Reprod Dev 29: pp. 385-391 CrossRef
- Wang, W, Day, B, Wu, G (2003) How does polyspermy happen in mammalian oocytes?. Microsc Res Tech 61: pp. 335-341 CrossRef
- Gil, M, Cuello, C, Parrilla, I, Vazquez, J, Roca, J, Martinez, E (2010) Advances in swine in vitro embryo production technologies. Reprod Domest Anim 45: pp. 40-48 CrossRef
- Macías García, B, González Fernández, L, Morrell, JM, Ortega Ferrusola, C, Tapia, JA, Rodriguez Martínez, H, Peña, FJ (2009) Single-layer centrifugation through colloid positively modifies the sperm subpopulation structure of frozen-thawed stallion spermatozoa. Reprod Domest Anim 44: pp. 523-526 CrossRef
- Macías García, B, Morrell, JM, Ortega-Ferrusola, C, González-Fernández, L, Tapia, JA, Rodríguez-Martínez, H, Peña, FJ (2009) Centrifugation on a single layer of colloid selects improved quality spermatozoa from frozen-thawed stallion semen. Anim Reprod Sci 114: pp. 193-202 CrossRef
- Roca, J, Rodríguez-Martínez, H, Vázquez, J, Bolarín, A, Hernández, M, Saravia, F, Wallgren, M, Martínez, E (2006) Strategies to improve the fertility of frozen-thawed boar semen for artificial insemination. Soc Reprod Fertil Suppl 62: pp. 261-275
- Vázquez, J, Parrilla, I, Roca, J, Gil, M, Cuello, C, Vázquez, J, Martínez, E (2009) Sex-sorting sperm by flow cytometry in pigs: issues and perspectives. Theriogenology 71: pp. 80-88 CrossRef
- Nakai, M, Kashiwazaski, N, Takizawa, A, Hayashi, Y, Nakatsukasa, E, Fuchimoto, D, Noguchi, J, Kaneko, H, Shino, M, Kikuchi, K (2003) Viable piglets generated from porcine oocytes matured in vitro and fertilized by intracytoplasmic sperm head injection. Biol Reprod 68: pp. 1003-1008 CrossRef
- Blomqvist, G, Persson, M, Wallgren, M, Wallgren, P, Morrell, J (2011) Removal of virus from boar semen spiked with porcine circovirus type 2. Anim Reprod Sci 126: pp. 108-114 CrossRef
- Single layer centrifugation-selected boar spermatozoa are capable of fertilization in vitro
- Open Access
- Available under Open Access This content is freely available online to anyone, anywhere at any time.
Acta Veterinaria Scandinavica
- Online Date
- March 2013
- Online ISSN
- BioMed Central
- Additional Links
- Sperm selection
- Author Affiliations
- 1. Department of Clinical Sciences, Division of Reproduction, Swedish University of Agricultural Sciences, P.O. Box 7054, Uppsala, SE 750 07, Sweden
- 2. Centre for Reproductive Biology in Uppsala, Swedish University of Agricultural Sciences, P.O. Box 7054, Uppsala, SE 750 07, Sweden