Surgical Oocyte Retrieval and the Developmental Potential of the Oocytes Derived from Prepubertal Calves

  • Yeh, S.P. (Department of Animal Science, National Chung Hsing University) ;
  • Ju, J.C. (Department of Animal Science, National Chung Hsing University) ;
  • Tseng, J.K. (Department of Animal Science, National Chung Hsing University) ;
  • Chou, P.C. (Department of Animal Science, National Chung Hsing University) ;
  • Chen, C.H. (Department of Animal Science, National Chung Hsing University) ;
  • Chih, T. (Department of Animal Science, National Chung Hsing University) ;
  • Fan, Y.K. (Department of Animal Science, National Chung Hsing University)
  • Received : 2003.05.21
  • Accepted : 2003.09.17
  • Published : 2004.02.01


The objectives of this study were to investigate the ovarian responsiveness of juvenile calves to exogenous gonadotropin treatments and to establish the oocyte retrieval technique for prepubertal heifers. Three 78-day-old calves were treated with 4 doses (40, 30, 30 and 30 mg) of FSH (Folltropin V) at 12 h interval up to 229 day-old. Surgical oocyte retrieval was performed 24 h after the last injection of FSH. Calves with good ovarian responses to FSH treatment had an average ovarian size of $5{\times}3$ cm compared to $3{\times}2$ cm in the less-responsive animals. Large variations were observed in the number of total follicles ($51{\pm}45$), aspirated follicles ($39{\pm}36$), oocytes recovered ($23{\pm}25$) and usable oocytes recovered ($11{\pm}19$) during 78 to 229 day-old. Oocytes derived from prepubertal calves had significantly lower maturation rate than those from cows (34 vs. 100%, p<0.05). Mean diameters of calf oocytes ($144{\pm}1{\mu}m$) and ooplasm ($110{\pm}1 {\mu}m$) were significantly lower than those of cows ($149{\pm}1$ and $25{\pm}1{\mu}m$, respectively). The diameter of the ooplasm also increased significantly after in vitro maturation (IVM) ($108{\pm}1$ vs. $112{\pm}1{\mu}m$). However, further studies are required to optimize the IVP system for the oocytes derived from prepubertal heifers.


Supported by : National Science Council


  1. Armstrong, D. T., B. J. Irvine, C. R. Earl, D. McLean and R. F. Seamark. 1994. Gonadotropin stimulation regimes for follicular aspiration and in vitro embryo production from calf oocytes. Theriogenology 42:1227-1236.
  2. Armstrong, D. T., P. Holm, B. J. Irvine, B. A. Petersen, R. B. Stubbings, D. McLean, G. F. Stevens and R. F. Seamark. 1992. Pregnancies and live births from in vitro fertilization of calf oocytes by laparoscopic follicular aspiration. Theriogenology 38:667-678.
  3. Gandolfi, F., E. Milanesi, P. Pocar, A. M. Luciano, T. A. L. Brevini, F. Acocella, A. Lauria and D. T. Armostrong. 1998. Comparative analysis of calf and cow oocytes during in vitro maturation. Mol Reprod Dev. 49:168-175.
  4. Kelly, J. C., J. R. Earl and D. T. Armstrong. 1997. Blastocyst production rate from different grades of oocytes collected from 2 to 3 month old calves. Theriogenology 47:291 (abstr).
  5. Majerus, V., R. De Roover, D. Etienne, S. Kaidi, A. Massip, F. Dessy and I. Donnay. 1999. Embryo production by ovum pick up in unstimulated calves before and after puberty. Theriogenology 52:1169-1179.
  6. Revel, F., P. Mermillod, N. Peynot, J. P. Renard and Y. Heyman. 1995. Low developmental capacity of in vitro matured and fertilized oocytes from calves compared with that of cows. J. Reprod. Fertil. 103:115-120.
  7. SAS. 1988. SAS Procedure Guide, Release 6.03 ed. SAS Institute Inc., Cary, NC, USA.
  8. Testart, J., G. Kann, J. Saumande and M. Thibier. 1977. Oestradiol-17 $\beta$, progesterone, FSH and LH in prepubertal calves induced to superovulate. J. Reprod. Fert. 51:329-336.
  9. Armstrong, D. T., P. J. Kotaras and C. R. Earl. 1997. Advances in production of embryos in vitro from juvenile and prepubertal oocytes from the calf and lamb. Reprod. Fertil. Dev. 9:333-339.
  10. Taneja, M., E. J. Peter, P. E. J. Bols, A. Van De Velde, J. C. Ju, D. Schreiber, M. W. Tripp, H. Levine, Y. Echelard, J. Riesen and X. Yang. 2000. Developmental competence of juvenile calf oocytes in vitro and in vivo: Influence of donor animal variation and repeated gonadotropin stimulation. Biol. Reprod. 62:206-213.
  11. Yang, X., C. Kubota, H. Suzuki, M. Taneja, P. E. J. Bols and G. A. Presicce. 1998. Control of oocyte maturation in cowsbiological factors. Theriogenology 49:471-482.
  12. Lohuis, M. M. 1995. Potential benefits of bovine embryomanipulation technologies to genetic improvement programs. Theriogenology 43:51-60.
  13. Martino, A., T. Mogas, M. J. Palomo and M. T. Paramio. 1995. In vitro maturation and fertilization of prepubertal goat oocytes. Theriogenology 43:473-485.
  14. Damiani, P., R. A. Fissore, J. B. Cibelli, J. M. Robl and R. T. Duby. 1995. Evaluation of cytoplasmic maturation of calf oocytes. Theriogenology 43:191 abstr.
  15. Duby, R. T., P. Damiani, C. R. Looney, C. R. Long, J. J. Balise and J. M. Robl. 1995. Cytological characterization of maturation and fertilization in prepubertal calf oocytes. Theriogenology 43:202 (abstr).
  16. Evans, A. C. O., G. P. Adams and N. C. Rawlings. 1994. Endocrine and ovarian follicular changes leading up to first ovulation in prepubertal heifers. J. Reprod. Fertil. 100:187-194.
  17. Liu, L., J. C. Ju and X. Yang. 1998. Parthengenetic development and protein pattern of newly matured bovine oocytes after chemical activation. Mol. Reprod. Dev. 49:298-307.
  18. Hashimoto, S., K. Saeki, Y. Nagano, N. Minami, M. Uamada and K. Utsumi. 1998. Effects of cumulus cell density during in vitro maturation on the developmental competence of bovine oocytes. Theriogenology 49:1451-1463.
  19. Khatir, H., P. Lonergan and P. Mermillod. 1998. Kinetics of nuclear maturation and protein profiles of oocytes from prepubertal and adult cattle during in vitro maturation. Theriogenology 50:917-929.
  20. Armstrong, D. T. 1993. Recent advances in superovulation of cattle. Theriogenology 39:7-24.
  21. Yang, X., G. A. Presecce, L. Moraghan, S. Jiang and R. H. Foote. 1994. Synergistic effect of ethanol and cycloheximide on activation of freshly matured bovine oocytes. Theriogenology 41:395-403.
  22. Duby, R. T., P. Damiani, C. R. Looney, R. A. Fissore and J. M. Robl. 1996. Prepubertal calves as oocyte donors: promises and problem. Theriogenology 45:121-131.
  23. Erickson, B. H. 1966. Development and senescence of the postnatal bovine ovary. J. Anim. Sci. 25:800-805.
  24. Ju, J. C., L. Liu, J. E. Parks and Yang X. 1998. Cytoskeletal reorganization of IVM-derived bovine oocytes after in vitro fertilization. Theriogenology 49:286 (abstr).
  25. Looney, C. R., P. Damiani, B. R. Lindsey, C. R. Long, C. L. Gonseth, D. L. Johnson and R. T. Duby. 1995. Use of prepubertal heifers as oocyte donors for IVF: effect of age and gonadotropin treatment. Theriogenology 43:269 abstr.
  26. Adams, G. P., A. C. O. Evans and N. C. Rawlings. 1994. Follicular waves and circulating gonadotrophins in 8-month-old prepubertal heifers. J. Reprod. Fertil. 100:27-33.
  27. Fair, T., P. Hyttel and T. Greve. 1995. Bovine oocyte diameter in relation to maturational competence and transcriptional activity. Mol. Reprod. Dev. 42:437-442.