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Developmental capacity of Antarctic minke whale (Balaenoptera bonaerensis) vitrified oocytes following in vitro maturation, and parthenogenetic activation or intracytoplasmic sperm injection

  • Takuma Fujihira (a1), Mariko Kobayashi (a1), Shinichi Hochi (a2), Masumi Hirabayashi (a3), Hajime Ishikawa (a4), Seiji Ohsumi (a4) and Yutaka Fukui (a1)...

The present study investigated the effects of the sexual maturity of oocyte donors on in vitro maturation (IVM) and the parthenogenetic developmental capacity of fresh minke whale oocytes. The effects of cytochalasin B (CB) pretreatment and two types of cryoprotectant solutions (ethylene glycol (EG) or ethylene glycol and dimethylsulfoxide (EG + DMSO)) on the in vitro maturation of vitrified immature whale oocytes were compared, and the developmental capacity of vitrified immature whale oocytes following IVM and intracytoplasmic sperm injection examined (ICSI). The maturation rate did not differ significantly with sexual maturity (adult, 60.9%; prepubertal, 53.1%), but the parthenogenetic activation rate of oocytes from adult donors (76.7%) was significantly higher (p < 0.05) than that of oocytes from prepubertal donors (46.4%). The maturation rates after vitrification and warming were not significantly different between the EG (22.2%) and EG + DMSO groups (30.2%), or between the CB-treated (30.4%) and non-CB-treated groups (27.3%). These results indicate that parthenogenetic activation of in vitro matured oocytes from adult minke whales was superior to that from prepubertal whales, but that the developmental capacity of the whale oocytes after parthenogenetic activation or ICSI was still low. The present study also showed that CB treatment before vitrification and two kinds of cryoprotectants did not improve the IVM rate following the vitrification of immature whale oocytes.

Corresponding author
All correspondence to: Y. Fukui. Fax: +81 155 495593. e-mail:
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Asada, M., Tetsuka, M., Ishikawa, H., Ohsumi, S. & Fukui, Y. (2001). Improvement on. in vitro maturation, fertilization and development of minke whale (Balaenoptera acutorostrata) oocytes. Theriogenology 56, 521–33.
Certin, Y. & Bastan, A. (2006). Cryopreservation of immature bovine oocytes by vitrification in straws. Anim. Reprod. Sci., 92, 2936.
Cha, K.Y., Chung, H.M., Lim, J.M., Ko, J.J., Han, S.Y., Choi, D.H. & Yoon, T.K. (2000). Freezing immature oocytes. Mol. Cell. Endocrinol. 169, 43–7.
Chang, S.C., Jones, J.D., Ellefson, R.D. & Ryan, R.J. (1976). The porcine ovarian follicle. I. selected chemical analysis of follicular fluid at different developmental stages. Biol. Reprod. 15, 321–8.
Dobrinsky, J.R. (2002). Advancements in cryopreservation of domestic animal embryos. Theriogenology 57, 285302.
Dobrinsky, J.R., Pursel, V.G., Long, C.R. & Johnson, L.A. (2000). Birth of piglets after transfer of embryos cryopreserved by cytoskeletal stabilization and vitrification. Biol. Reprod. 62, 564–70.
El Mouatassim, S., Guérin, P., Ménézo, Y. (1999). Expression of genes encoding antioxidant enzymes in human and mouse oocytes during the final stages of maturation. Mol. Hum. Reprod. 5, 720–5.
Fujihira, T., Kishida, R. & Fukui, Y. (2004). Developmental capacity of vitrified immature porcine oocytes following ICSI: effects of cytochalasin B and cryoprotectants. Cryobiology 49, 286–90.
Fukui, Y., Mogoe, T., Terawaki, Y., Ishikawa, H., Fujise, Y. & Ohsumi, S. (1995). Relationship between physiological status and serum constituent values in minke whales (Balaenoptera acutorostrata). J. Reprod. Dev. 41, 203–8.
Fukui, Y., Mogoe, T., Ishikawa, H. & Ohsumi, S. (1997 a). Factors affecting in vitro maturation of minke whale (Balaenoptera acutorostrata) follicular oocytes. Biol. Reprod. 56, 523–8.
Fukui, Y., Mogoe, T., Ishikawa, H. & Ohsumi, S. (1997 b). In vitro fertilization of in vitro matured minke whale (Balaenoptera acutorostrata) follicular oocytes. Mar. Mamm. Sci. 13, 395404.
Funahashi, H., Cantley, T.C., Stumpf, T.T., Terlouw, S.L. & Day, B.N. (1994). Use of low-salt culture medium for in vitro maturation of porcine oocytes is associated with elevated oocyte glutathione levels and enhanced male pronuclear formation after in vitro fertilization. Biol. Reprod. 51, 633–9.
Iwata, H., Hashimoto, S., Ohota, M., Kimura, K., Shibano, K. & Miyake, M. (2004). Effects of follicle size and electrolytes and glucose in maturation medium on nuclear maturation and developmental competence of bovine oocytes. Reproduction 127, 159–64.
Isachenko, V., Soler, C., Isachenko, E., Perez-Sanchez, F. & Grishchenko, V. (1998). Vitrification of immature porcine oocytes: effects of lipid droplets, temperature, cytoskeleton, and addition and removal of cryoprotectant. Cryobiology 36, 250–3.
Iwayama, H., Hochi, S., Kato, M., Hirabayashi, M., Kuwayama, M., Ishikawa, H., Ohsumi, S. & Fukui, Y. (2004). Effects of cryodevice type and donors’ sexual maturity on vitrification of minke whale (Balaenoptera bonaerensis) oocytes at germinal vesicle stage. Zygote 12, 333–8.
Iwayama, H., Ishikawa, H., Ohsumi, S. & Fukui, Y. (2005). Attempt at in vitro maturation of minke whale (Balaenoptera bonaerensis) oocytes using a portable CO2 incubator. J. Reprod. Dev. 51, 6975.
Katayama, K.P., Stehlik, J., Kuwayama, M., Kato, O. & Stehlik, E. (2003). High survival rate of vitrified human oocytes results in clinical pregnancy. Fertil. Steril. 80, 223–4.
Kishida, R., Lee, E.S. & Fukui, Y. (2004). In vitro maturation of porcine oocytes using a defined medium and developmental capacity after intracytoplasmic sperm injection. Theriogenology 62, 1663–76.
Marchal, R., Feugang, J.M., Perreau, C., Venturi, E., Terqui, M., Mermillod, P. (2001). Meiotic and developmental competence of prepubertal and adult swine oocytes. Theriogenology 56, 1729.
O'Brien, J.K., Dwarte, D., Ryan, J.P., Maxwell, W.M.C. & Evans, G. (1996). Developmental capacity, energy metabolism and ultrastructure of mature oocytes from prepubertal and adult sheep. Reprod. Fertil. Dev. 8, 1029–37.
Revel, F., Mermillod, P., Peynot, N., Renard, J.P. & Heyman, Y. (1995). Low developmental capacity of. in vitro matured and fertilized oocytes from calves compared with that of cows. J. Reprod. Fertil. 103, 115–20.
Roblero, L., Biggers, J.D. & Lechene, C.P. (1976). Electron probe analysis of the elemental microenvironment of oviducal mouse embryos. J. Reprod. Fertil. 46, 431–4.
Suzuki, M., Misumi, K., Ozawa, M., Noguchi, J., Kaneko, H., Ohnuma, K., Fuchimoto, D.I., Onishi, A., Iwamoto, M., Saito, N., Nagai, T. & Kikuchi, K. (2006). Successful piglet production by IVF of oocytes matured in vitro using NCSU-37 supplemented with fetal bovine serum. Theriogenology, 65, 374–86.
Vieira, A.D., Mezzalira, A., Barbieri, D.P., Lehmkuhl, R.C., Rubin, M.I. & Vajta, G. (2002). Calves born after open pulled straw vitrification of immature bovine oocytes. Cryobiology 45, 91–4.
Wang, W.H., Machaty, Z., Abeydeera, L.R., Prather, R.S. & Day, B.N. (1998). Parthenogenetic activation of pig oocytes with calcium ionophore and the block to sperm penetration after activation. Biol. Reprod. 58, 1357–66.
Wise, T. (1987). Biochemical analysis of bovine follicular fluid: albumin, total protein, lysosomal enzymes, ions, steroids and ascorbic acid content in relation to follicular size, rank, atresia classification and day of estrous cycle. J. Anim. Sci. 64, 1153–69.
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