Japanese society of Ova Research

Abstract

Vol.37 No.1

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Successful vitrification of rat embryos using a carboxylated ε-poly-L-lysine
JMOR, 37(1) 31-35, 2020
Maki Kamoshita1# Narumi Nakano2# Katsuyoshi Fujiwara1 Ayumi Suyama2 Kazuaki Matsumura3 Suong-Hyu Hyon4 Junya Ito1,2* Naomi Kashiwazaki1,2 
1Graduate School of Veterinary Science, Azabu University, 1-17-71 Fuchinobe, Chuo-ku, Sagamihara, Kanagawa 252-5201, Japan, 2Laboratory of Animal Reproduction, School of Veterinary Medicine, Azabu University, 1-17-71 Fuchinobe, Chuo-ku, Sagamihara, Kanagawa 252-5201, Japan, 3School of Materials Science, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi, Ishikawa 923-1292, Japan, 4Lab, Joint Faculty of Veterinary Medicine, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan

Recently, production of genetically modified animals using a genome editing system such as CRISPR/Cas9 has been useful for modifying various species including rats. Such genetically modified rats can be preserved as germ cells to reduce the costs of maintenance of live animals. Vitrification of preimplantation embryos is valuable for many researchers. However, it is known that rat embryos are sensitive to physiological and physical stress and further improvement of the vitrification protocol is required. In this study, we examined whether a new cryoprotective agent (CPA), carboxylated ε-poly-L-lysine (COOH-PLL), is suitable for vitrification of rat embryos from the pronuclear to blastocyst stages in a minimal volume cooling protocol. Our results show that most of the embryos vitrified with ethylene glycol and COOH-PLL survived and developed to the blastocyst. After the transfer of vitrified embryos, the survival rate to term was similar to that of fresh embryos. We conclude that our vitrification protocol is suitable for rat embryos.

Keywords: Rat  Embryo  Vitrification 
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