Efecto de la heparina y de la concentración espermática sobre el porcentaje de fertilización de oocitos bovinos in vitro

Authors

  • Roger D. Salgado
  • Clara C. Rugeles
  • Jaime Alvarez

DOI:

https://doi.org/10.17533/udea.rccp.324000

Keywords:

bovino, fertilización in vitro, heparina, swim up

Abstract

ResumenEl objetivo de este estudio fue evaluar el efecto de diferentes concentraciones de heparina y de semen en el medio de fertilización sobre el porcentaje de oocitos bovinos fertilizados in vitro. En el experimento uno, los oocitos obtenidos de ovarios de matadero fueron madurados in vitro y distribuidos al azar en seis tratamientos para fertilización in vitro, de acuerdo a las concentraciones de heparina (2,4, 8, 10, 16, 20 µg/ml). En el experimento dos, una vez seleccionada la concentración de heparina en experimento 1, se evaluaron tres dosis de espermatozoides en el medio de fertilización (0.5x106 espermatozoides/ml, 1x106 espermatozoides/ml y 2x106 espermatozoides/ml). En cada experimento se utilizó un diseño completamente al azar y los datos fueron analizados por ANAVA. La concentración de heparina tuvo efecto (p < 0.05) en el porcentaje de oocitos fertilizados encontrándose los mejores resultados en la concentración de 10 µg/ml, con una tasa de penetración del 67%.  La concentración espermática afecta la fertilización, encontrándose la mayor  proporción de oocitos con fertilización normal (74%) al utilizar concentración 2x106 espermatozoides/ml. Sin embargo,  no fue significativamente diferente a la concentración de 1x106 (p > 0.05). Los resultados indican que la tasa de fertilización guarda relación directa con la dosis de heparina y la concentración espermática.

SummaryThe objective of this study was to evaluate the effect of different heparin concentrations and of the semen in the percentage of bovine oocytes in fertilization medium. In the experiment 1, the obtained oocytes of slaughterhouse ovaries were matured in vitro and distributed at random in six treatments for fertilization in vitro, according to the heparin concentration (2, 4, 8, 10, 16, 20 µg/ml). In the experiment 2, once selected the heparin concentration from experiment 1, three sperms doses were evaluated in fertilization medium (0.5x106espermatozoas/ml, 1x06espermatozoas/ml and 2x06espermatozoas/ml). In each experiment totally randomized design was used and data were analyzed by ANAVA. The concentration of heparin (p < 0.05) affected the percentage of fertilized oocytes being the best results in the concentration of 10 µg/ml, with a rate of penetration of 67%. The spermatic concentration affects the fertilization, the biggest oocytes proportion with normal fertilization (74%) when using concentration 2x106spermatozoas/ml. However, it was not significantly different to the concentration of 1x 106 (p > 0.05). The results indicate that the fertilization rate keeps direct relationship with the heparine dose and the spermatic concentration.

 

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References

Camargo LS, Sá W, Ferreira AM, Viana JH. Efeito de sistemade cultivo, célula somática e soro em co-cultura sobre odesenvolvimento de embrioes bovinos fecundados in vitro.Arq Bras Med Vet Zootec 2001; 23:78-83.

Camargo LS, Sá W, Ferreira AM, Viana JH, Freitas C.Concentracão espermática na fecundacão in vitro, com semende touro da raca Guzará. Arq Bras Med Vet Zootec 2000;52:59-64.

Chacon LJ, Estrada JL. Evaluación de diferentes compuestosen la estandarización de la técnica de fertilización in vitro enbovinos. Tesis para Msc. Universidad Nacional Colombia(1997).

Eyestone W, First N. Co-culture of early cattle embryos tothe blastocyst stage with oviductal tissue or in conditionedmedium. J Reprod Fertil 1989; 85:715-720.

Ferry L, Mermillod P, Massip A, Dessy F. Bovine embryosculture in serum-poor oviduct-conditioned medium needcooperation to reach the blastocist stage. Theriogenology1994; 42: 445-453.

Hernández Ledezma JJ, Villanueva C, Sikes JD, RobertsRM. Effects of CZB versus mediuim 199 and of conditioningcultura media with either bovine oviductal epithelial cells orbuffalo rat liver cells on the development of bovine zygotesderived by in vitro maduretion-in vitro fertilizationprocedures. Theriogenology 1993; 39:1267-1277.

Kim BK, Lee SC, Lee KS, Lee BK, Kim JH. Effect ofmedium milieu on sperm penetration and pronuclearformation of bovine oocitos matured in vitro.Theriogenology.2002; 57:2093 -104.

Kurtu JM, Ambrose JD, Rajamahendran R. Cleavage rata ofbovine oocytes in-vitro is affected by bulls but not spermconcentrations. Theriogenoloy 1996; 45:257 – 265.

Leven K, Clay A, Burnley A, Benjamin G. Production ofviable bovine blastocysts in defined in vitro conditions.Biology of reproduction 1995; 52:1410 – 1417.

Lu KH, Seidel GE Jr. Effects of heparin and spermconcentration on cleavage and blastocyst development ratesof bovine oocytes inseminated with flow cytometrically-sorted sperm. Theriogenology 2004; 62:819-30.

Niwa, K, Ohgoda, O. Synergistic effect of caffein and heparinon in-vitro fertilization of cattle oocytes matured in culture.Theriogenology 1988; 30:733-741.

Parrish JJ, Susko-Parrish, First NL. Effect of heparin andchondroitin sulfate on the acrosome reaction and fertilityof bovine sperm in vitro. Theriogenology. 1985; 24: 537-549.

Parrish JJ, Susko-Parrish JL, Leibfried Rufledge ML, CristerEJ Eyestone EH, First NL. Bovine in-vitro fertilization withfrozen-thawed semen. Theriogenoloy 1986; 25:591-600.

Parrish JJ, Susko-Parrish MA, Winer A, First NL.Capacitation of bovine sperm by heparin. Biol Reprod 1988;38:1171-1180.

Parrish JJ, Susko Parrish JL, Handrow RR, Sims MM andFirst NL. Capacitation of bovine spermatozoa by oviductfluid. Biol Reprod 1989; 40:1020-1025.

Pinyopummintr T. and Bavister B. In vitro- matured/invitro- fertilized bovine oocytes can develop into morulae/blastocists in chemically defined, protein- free culture media.Biol Reprod 1991; 45: 736-742.

Rieger D, Grisart B, Semple E, Van Langendonckt A,Betteridge KJ, et al. Comparison of the effects of oviductalcell co-culture and oviductal cell-conditioned medium on thedevelopment and metabolic activity of cattle embryos. JReprod Fertil 1995; 105: 91-98.

Saeki K, Nagao Y, Hoshi M and Nagai M. Effect of heparin,sperm concentration and bull variation on in vitro fertilizationof bovine oocytes in a protein-free medium. Theriogenology1995; 43:751-759.

Shamsuddin M, Niwa K, Larsson B, Rodriguez-MartinezH. In vitro maturation and fertilization of bovine oocytes.Reprod Dom Anim 1996; 31:613-622.

Xu KP, Greve T. A detailed analysis of early events duringin-vitro fertilization of bovine folicular oocytes. J ReprodFertil 1988; 82:127-134.

Van Inzen WG, Van Stekelenburg-Hamers AE, Weima SM,Kruip TA, Bevers MM, et al. Culture of bovine embryos tothe blastocist stage using buffalo rat liver (BRL) cells.Theriogenology 1995; 43:723-738.

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Published

2016-07-19

How to Cite

Salgado, R. D., Rugeles, C. C., & Alvarez, J. (2016). Efecto de la heparina y de la concentración espermática sobre el porcentaje de fertilización de oocitos bovinos in vitro. Revista Colombiana De Ciencias Pecuarias, 18(2), 5. https://doi.org/10.17533/udea.rccp.324000

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