Retrieval of Porcine Ovarian Follicles by Different Methods

  • Choi, Moon Hwan (Gamete and Stem Cell Biotechnology, Seoul National University) ;
  • Gong, Seung Pyo (Gamete and Stem Cell Biotechnology, Seoul National University) ;
  • Lim, Jeong Mook (Gamete and Stem Cell Biotechnology, Seoul National University)
  • 투고 : 2006.12.16
  • 심사 : 2007.08.23
  • 발행 : 2008.03.01


A series of studies has been conducted to establish a base infrastructure for an ovarian follicle culture system in the porcine and this study was designed to develop an effective retrieval protocol of preantral follicles. Five different methods using collagenase type I (A) or IV (B, C1, C2 and C3), which employed different treatment durations and/or conditions, were employed and sliced ovarian tissue of prepubertal gilts was provided for the retrieval. A significant increase in total number of follicles retrieved was detected when collagenase IV (methods B or C) was used. In total, more ovarian follicles were retrieved by method B undertaking agitation and method C2 without the agitation than method C1 and C3, while the number of preantral follicles collected was the largest in method B. Neither incubation in 5% $CO_2$ in air atmosphere instead of the agitation nor increased duration of enzymatic treatment up to 120 minutes improved the efficiency of follicle retrieval. There were no differences in the number of follicles retrieved from intact ovaries and from used ovaries for oocyte collection. These results demonstrate the collagenase IV treatment with agitation is effective for retrieving porcine preantral follicles from the ovaries.


Porcine;Ovarian Follicle;Retrieval;Oocytes


연구 과제 주관 기관 : Rural Development Administration, Republic of Korea


  1. Choi, J. K., J. H. Lee, S. T. Lee, M. H. Choi, S. P. Gong, E. J. Lee and J. M. Lim. 2007. Developmental competence of intrafollicular oocytes derived from preantral follicle culture with different protocols after parthenogenetic activation. Asian-Aust. J. Anim. Sci. 20:1190-1195.
  2. Cortvrindt, R. G., Y. Hu, J. Liu and J. E. J. Smitz. 1998. Timed analysis of the nuclear maturation of oocytes in early preantral mouse follicle culture supplemented with recombinant gonadotropin. Fertil. Steril. 70:1114-1125.
  3. Akshey, Y. S., P. Palta, R. S. Manik, Vivekananad and M. S. Chauhan. 2005. Effect of removal of follicles through repeated transvaginal follicle aspiration on subsequent follicular populations in murrah buffalo (Bubalus bubalis). Asian-Aust. J. Anim. Sci. 18:632-636.
  4. Mao, J., G. Wu, M. F. Smith, T. C. McCauley, T. C. Cantley, R. S. Prather, B. A. Didion and B. N. Day. 2002. Effects of culture medium, serum type, and various concentrations of folliclestimulating hormone on porcine preantral follicular development and antrum formation in vitro. Biol. Reprod. 67: 1197-1203.
  5. McShane, P., R. Sutton, D. W. Gray and P. J. Morris. 1989. Protease activity in pancreatic islet isolation by enzymatic digestion. Diabetes. 1:126-128.
  6. Lenie, S., R. Cortvrindt, T. Adriaenssens and J. Smitz. 2004. A reproductive two-step culture system for isolated primary mouse ovarian follicles as single functional units. Biol. Repod. 71:1730-1738.
  7. Eppig, J. J. and M. O'Brein. 1996. Development in vitro of mouse oocytes from primordial follicles. Biol. Reprod. 54:197-207.
  8. Hirao, Y., T. Nagai, M. Kubo, M. Miyano and S. Kato. 1994. In vitro growth and maturation of pig oocytes. J. Reprod. Fertil. 100:333-339.
  9. Lee, S. T., M. H. Choi, E. J. Lee, S. P. Gong, M. Jang, S. H. Park, H. Jee, D. Y. Kim, J. Y. Han and J. M. Lim. 2007. Establishment of autologous embryonic stem cells derived from preantral follicle culture and oocyte parthenogenesis. Fertil. Steril. in press.
  10. Donnelly, K. and E. E. Telfer. 1994 Growth of porcine preantral ovarian follicles in different culture systems. J. Reprod. Fertil. Abstract Series. 13:abstract 76.
  11. Eppig, J. J. and A. C. Schroeder. 1989. Capacity of mouse oocytes from preantral follicles to undergo embryogenesis and development to live young after, growth, maturation, and fertilization in vitro. Biol. Reprod. 41:268-276.
  12. Xu, X. M., J. L. Hua, W. W. Jia, W. Huang, C. R. Yang and Z. Y. Dou. 2007. Parthenogenetic activation of porcine oocytes and isolation of embryonic stem cells-like derived from parthenogenetic blastocysts. Asian-Aust. J. Anim. Sci. 20:1510-1516.
  13. Wu, M. F., W. T. Huang, C. Tsay, H. F. Hsu, B. T. Liu, C. M. Chiou, S. C. Yen, S. P. Cheng and J. C. Ju. 2002. The stagedependent inhibitory effect of porcine follicular cells on the development of preantral follicles. Anim. Reprod. Sci. 73:73- 88.
  14. Wu, J., B. R. Emery and D. T. Carrell. 2001. In vitro growth, maturation, fertilization, and embryonic development of oocytes from porcine preantral follicles. Biol. Reprod. 64:375- 381.
  15. Telfer, E. E. 1996. The development of methods for isolation and culture of preantral follicles from bovine and porcine ovaries. Theriogenol. 45:101-110.
  16. Smitz, J. E. and R. G. Cortvrindt. 2001. The earliest stages of folliculogenesis in vitro. Reprod. 123:185-202.
  17. Choi, M. H., J. H. Oh, T. M. Kim, J. Y. Han and J. M. Lim. 2006. Morphological criteria of bovine ovaries for predicting retrieval efficiency of preantral follicles. Asian-Aust. J. Anim. Sci. 19:1711-1715.
  18. Denning, C. and H. Priddle. 2003. New frontiers in gene targeting and cloning; success, application and challenges in domestic animals and human embryonic stem cells. Reprod. 126:1-11.
  19. Li, M., D. Zhang, Y. Hou, L. Jiao, X. Zheng and W. H. Wang. 2003. Isolation and culture of embryonic stem cells from porcine blastocysts. Mol. Reprod. Dev. 65:429-434.
  20. Morbeck, D. E., K. L. Esbenshade, W. L. Flowers and J. H. Britt. 1994. Kinetics of follicle growth in the prepubertal gilt. Biol Reprod. 47:485-491.
  21. Shang, J. H., R. X. Xu, X. D. Jiang, Y. X. Zou, L. S. Qin, Y. Q. Cai, Z. J. Yang, X. Zheng and S. Cui. 2007. Somatic cell nuclear transfer of oocytes aspirated from postovulatory ovarian follicles of superovulated rabbits. Asian-Aust. J. Anim. Sci. 20:1354-1360.
  22. Yang, X. Y., H. Li, W. Y. Huang, S. Z. Huang and Y. Zeng. 2005. Comparison of two different schemes of once-weekly ovum pick up in dairy heifers. Asian-Aust. J. Anim. Sci. 18:314-319.

피인용 문헌

  1. Isolation and Culture of Preantral Follicles for Retrieving Oocytes for the Embryo Production: Present Status in Domestic Animals vol.47, pp.3, 2011,