Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
권호 표지

In Vitro Maturation of Porcine Oocytes in a Dry Incubator without $CO_2$ Gas Supplement

  • Park, Kwang-Wook (Dept. of Animal Science & Technology, Sunchon National University)
  • Received : 2012.09.07
  • Accepted : 2012.09.08
  • Published : 2012.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

The present study was conducted to develop a simple method for porcine oocyte maturation without $CO_2$ regulation. In experiment 1, we evaluated that the effect of $CO_2$ non-supplement on porcine oocyte maturation. Cumulusoocyte complexes (COCs) were collected from 2~6 mm follicles and divided into three groups (Control, tube-$CO_2$, and tube-non-$CO_2$). For control, COCs were cultured in 4-well multidish in a $CO_2$ incubator. For tube-$CO_2$, COCs were cultured in a round-bottom tube in a $CO_2$ incubator, and for tube-non-$CO_2$, COCs were cultured in a round-bottom tube sealed tightly without $CO_2$ supplement in a dry incubator. The proportion of oocytes reached to metaphase II (M-II) was not significantly different among three groups (87.9% to 91.4%). In experiment 2, we evaluated the effect of $CO_2$ non-supplement during oocyte maturation on development of embryos. Oocytes with a polar body were divided into two groups (Control and tube-non-$CO_2$) and applied 1.1 kV/cm or 1.2 kV/cm voltages for parthenogenetic activation. After activation, embryos were cultured for 6 days and examined the development. The proportion of embryos cleaved was not significantly different among treatment (86.3% to 91.5%). The proportion of embryo reached to blastocyst stage was not significantly different among treatment (13.9% to 25.2%). The cell number of blastocysts was not significantly different among treatment (29.0 to 32.4). In conclusion, oocytes cultured in a dry incubator without $CO_2$ supplement have enough competence to development after parthenogenetic activation. These results would be useful for transporting oocytes or embryos a long distance.

Keywords

References

  1. Abeydeera LR, Wang WH, Cantley TC, Rieke A, Murphy CN, Prather RS, and Day BN (2000): Development and viability of pig oocytes matured in a protein- free medium containing epidermal growth factor. Theriogenology 54:787-797. https://doi.org/10.1016/S0093-691X(00)00390-3
  2. Bavister BD (1995): Culture of preimplantation embryos: facts and artifacts. Hum Reprod Update 1:91-148. https://doi.org/10.1093/humupd/1.2.91
  3. Byrd SR, Flores-Foxworth G, Applewhite AA, Westhusin ME (1997): In vitro maturation of ovine oocytes in a portable incubator. Theriogenology 47:857-864. https://doi.org/10.1016/S0093-691X(97)00041-1
  4. Chen N, Liow SL, Yip WY, Tan LG, Ng SC (2005): Influence of cysteamine supplementation and culture in portable dry-incubator on the in vitro maturation, fertilization and subsequent development of mouse oocytes. Theriogenology 63:2300-2310. https://doi.org/10.1016/j.theriogenology.2004.10.007
  5. Fujii A, Kaedei Y, Tanihara F, Ito A, Hanatate K, Kikuchi K, Nagai T, Otoi T (2010): In vitro maturation and development of porcine oocytes cultured in a straw or dish using a portable incubator with a $CO_2$ chamber. Reprod Domest Anim 45:619-624.
  6. Gandolfi F, Moor RM (1987): Stimulation of early embryonic development in the sheep by co-culture with oviductal epithelial cells. J Reprod Fertil 81:23-28. https://doi.org/10.1530/jrf.0.0810023
  7. Geshi M, Yonai M, Sakaguchi M, Nagai T (1999): Improvement of in vitro co-culture systems for bovine embryos using a low concentration of carbon dioxide and medium supplement with b-mercaptoethanol. Theriogenology 51:551-558. https://doi.org/10.1016/S0093-691X(99)00009-6
  8. Iwayama H, Ishikawa H, Ohsumi S, Fukui Y (2005): Attempt at in vitro maturation of minke whale (Balaenoptera Bonaerensis) oocytes using a portable $CO_2$ incubator. J Reprod Dev 51:69-75. https://doi.org/10.1262/jrd.51.69
  9. Kane MT (1972): Energy substrates and culture of single cell rabbit ova to blastocyst. Nature 238:468- 469. https://doi.org/10.1038/238468a0
  10. Karja NW, Wongsrikeao P, Murakami M, Agung B, Fahrudin M, Nagai T, Otoi T (2004): Effects of oxygen tension on the development and quality of porcine in vitro fertilized embryos. Theriogenology 62: 1585-1595. https://doi.org/10.1016/j.theriogenology.2004.03.012
  11. Krisher RL, Gibbons JR, Gwazdauskas FC (1998): Effectiveness of Menuzo's B2 medium with buffalo rat liver cells for development of in vitro matured/ in vitro fertilized bovine oocytes. J Assist Reprod Genet 15:50-53. https://doi.org/10.1023/A:1022582422932
  12. Li R, Murphy CN, Spate L, Wax D, Isom C, Rieke A, Walters EM, Samuel M, Prather RS (2009): Production of piglets after cryopreservation of embryos using a centrifugation-based method for delipation without micromanipulation. Biol Reprod 80:563-571. https://doi.org/10.1095/biolreprod.108.073387
  13. Love LB, Choi YH, Love CC, Varner DD, Hinrichs K (2003): Effect of ovary storage and oocyte transport method on maturation rate of horse oocytes. Theriogenology 59:765-774. https://doi.org/10.1016/S0093-691X(02)01133-0
  14. Menino AR, Wright RW (1982): Development of onecell porcine embryos in two culture systems. J Anim Sci 54:583-588. https://doi.org/10.2527/jas1982.543583x
  15. Nagai T (2001): The improvement of in vitro maturation systems for bovine and porcine oocytes. Theriogenology 55:1291-1301. https://doi.org/10.1016/S0093-691X(01)00483-6
  16. Niemann H, Rath D (2001): Progress in reproductive biotechnology in swine. Theriogenology 56:1291- 1304. https://doi.org/10.1016/S0093-691X(01)00630-6
  17. Ozawa M, Nagai T, Kaneko H, Noguchi J, Ohnuma K, Kikuchi K (2006): Successful pig embryonic development in vitro outside a $CO_2$ gas-regulated incubator: effects of pH and osmolality. Theriogenology 65:860-869. https://doi.org/10.1016/j.theriogenology.2005.06.012
  18. Petters RM, Wells KD (1993): Culture of pig embryos. J Reprod Fertil Suppl 48:61-73.
  19. Suzuki T, Sumantri C, Khan NH, Murakami M, Saha S (1999): Development of a simple, portable carbon dioxide incubator for in vitro production of bovine embryos. Anim Reprod Sci 54:149-157. https://doi.org/10.1016/S0378-4320(98)00134-1
  20. Varisanga MD, Dong YJ, Mtango NR, Fahrudin M, Ni Wayan KK, Suzuki T (2000): Bovine blastocysts obtained from reconstructed cytoplast and karyoplasts using a simple portable $CO_2$ incubator. Cloning 2: 167-173. https://doi.org/10.1089/152045500454726
  21. Whitten WK (1956): Culture of tubal mouse ova. Nature 177:96.
  22. Whitten WK, Biggers JD (1968): Complete development in vitro of the pre-implantation stages of the mouse in a simple chemically defined medium. J Reprod Fertil 17:399-401. https://doi.org/10.1530/jrf.0.0170399
  23. Xu KP, Greve T, Callesen H (1987): Pregnancy resulting from cattle oocytes matured and fertilized in vitro. J Reprod Fertil 81:501-504. https://doi.org/10.1530/jrf.0.0810501