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10 October 2012 Hollow Fiber Vitrification Provides a Novel Method for Cryopreserving In Vitro Maturation/Fertilization-Derived Porcine Embryos
Miki Maehara, Hitomi Matsunari, Kasumi Honda, Kazuaki Nakano, Yasuhiro Takeuchi, Takahiro Kanai, Taisuke Matsuda, Yukina Matsumura, Yui Hagiwara, Norihisa Sasayama, Akio Shirasu, Masashi Takahashi, Masahito Watanabe, Kazuhiro Umeyama, Yutaka Hanazono, Hiroshi Nagashima
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Abstract

In vitro matured (IVM) oocytes have been used to create genetically modified pigs for various biomedical purposes. However, porcine embryos derived from IVM oocytes are very cryosensitive. Developing improved cryopreservation methods would facilitate the production of genetically modified pigs and also accelerate the conservation of genetic resources. We recently developed a novel hollow fiber vitrification (HFV) method; the present study was initiated to determine whether this new method permits the cryopreservation of IVM oocyte-derived porcine embryos. Embryos were created from the in vitro fertilization of IVM oocytes with frozen-thawed sperm derived from a transgenic pig carrying a humanized Kusabira-Orange (huKO) gene. Morula-stage embryos were assigned to vitrification and nonvitrification groups to compare their in vitro and in vivo developmental abilities. Vitrified morulae developed to the blastocyst stage at a rate similar to that of nonvitrified embryos (66/85, 77.6% vs. 67/84, 79.8%). Eighty-eight blastocysts that developed from vitrified morulae were transferred into the uteri of three recipient gilts. All three became pregnant and produced a total of 17 piglets (19.3%). This piglet production was slightly lower, albeit not significantly, than that of the nonvitrification group (27/88, 30.7%). Approximately half of the piglets in the vitrification (10/17, 58.8%) and nonvitrification (15/27, 55.6%) groups were transgenic. There was no significant difference in the growth rates among the piglets in the two groups. These results indicate that the HFV method is an extremely effective method for preserving cryosensitive embryos such as porcine in vitro maturation/fertilization-derived morulae.

Miki Maehara, Hitomi Matsunari, Kasumi Honda, Kazuaki Nakano, Yasuhiro Takeuchi, Takahiro Kanai, Taisuke Matsuda, Yukina Matsumura, Yui Hagiwara, Norihisa Sasayama, Akio Shirasu, Masashi Takahashi, Masahito Watanabe, Kazuhiro Umeyama, Yutaka Hanazono, and Hiroshi Nagashima "Hollow Fiber Vitrification Provides a Novel Method for Cryopreserving In Vitro Maturation/Fertilization-Derived Porcine Embryos," Biology of Reproduction 87(6), (10 October 2012). https://doi.org/10.1095/biolreprod.112.100339
Received: 5 March 2012; Accepted: 1 October 2012; Published: 10 October 2012
KEYWORDS
cryopreservation
hollow fiber vitrification
in vitro produced porcine embryo
IVM/IVF
pig
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