IN VITRO MATURATION OF BOVINE OOCYTES FOR IN VITRO FERTILIZATION
Keywords:bovine, oocyte, in vitro fertilization, maturation, TCM 199, cultivation
The objective of the present study was to asses the quality of various cultivation media used for the maturation of bovine oocytes that are prepared for IVF. Upon collection from slaughtered bovine ovaries and after morphological evaluation, a total number of 513 viable oocytes have been selected for cultivation, being divided into 3 batches, 171 oocytes / batch. The oocytes belonging to batch 1 were cultivated in TCM 199 NaHCO3 + 10% FCS + FSH 20 μl/ml. The oocytes belonging to batch 2 were cultivated in TCM 199 NaHCO3 + 10% FCS + HCG 2.3 x 103 UI/ml + FSH 8 μl/ml + pyruvate 0.25 mM + 17β estradiol 1 μl/ml. The oocytes belonging to batch 3 were cultivated in TCM 199 NaHCO3 + 10% FCS + 17β estradiol 1 μl/ml + FSH 20 μl/ml. The cultivation conditions, for all three batches, were: 24 hours at 39°C, 5% CO2. Spermatozoa have been prepared using the Percoll method and IVF of the matured oocytes has been performed. Embryonic development has been assessed 72 hours and then up to 10 days after IVF. The results showed the superior quality of the oocytes belonging to batch 2 and matured using TCM 199 NaHCO3 + 10% FCS + HCG 2.3x103 UI/ml + FSH 8 μl/ml + pyruvate 0.25 mM + 17β estradiol 1 μl/ml, as their use for IVF yielded the highest number of viable embryos.
Arlotto T., J.L. Schwartz, N.L. First, M.L. Leibfried-Rutledge, 1996, Aspects of follicle and oocyte stage that affect in vitro maturation and development of bovine oocytes. Theriogenology 45, 943-956.
Fabbri, R., E. Porcu, T. Marsella, 2000, Technical aspects of oocytes cryopreservation. Molecular Cell and Endocrinology 169, 39-42.
Groza I, L. Harceaga, D. Moise, L. Bogdan, I. Morar, O. Sotoc, S. Ciupe,1996, Cercetări privind fecundarea “in vitro” a ovulelor de vaca. Simpozion – Lucrări Ştiinţifice de Medicină Veterinară, Timişoara, p. 271.
Groza I. (coordonator), L. Bogdan, R. Cătană, M. Cenariu, Simona Ciupe, D. Ciupercescu, I. Morar, M. Muntean, Al.R. Pop, Brînduşa Stegeran, Ginecologie, Andrologie şi Obstetrică Veterinară – Compendiu, Ed. Academiei Române, Bucureşti, 2006.
Holm, P., P.J. Booth, M.H. Schmidt, T. Greve, H. Callesen, 1999, High bovine blastocyst development in a static in vitro production system using SOFaa medium supplemented with sodium citrate and myo-inositol with or without serum-proteins. Theriogenology 52: 683-700.
Ito, K., M. Takahashi, K. Kawahata, T. Goto, J. Takahashi, Y. Yasuda, 1998, Supplementation effect of early pregnancy factor-positive serum into bovine in vitro fertilization culture medium. Animal Journal of Immunology 39(6), 356-361.
Sirad, M.A., R.D. Lambert, P. Guay, D.P. Menard, M. Bedoya, 1985, In vivo and in vitro development of in vitro fertilized bovine follicular oocytes obtained by laparoscopy. Theriogenology 23, 230.
Yang, X., C. Kubota, H. Suzuki, M. Taneja, P.E.J. Bols, G.A. Presicce, 1998, Control of oocyes maturation in cows-biological factors. Theriogenology 49, 471-482.
Younis, A.I., B.G. Brakett, R.A. Fayrer-Hosken, 1998, Influence of serum and hormones on bovine oocyte maturation and fertilization in vitro. Gamete Research 23, 189-209.
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