Successful vitrification of pronuclear-stage pig embryos with a novel cryoprotective agent, carboxylated ε-poly-L-lysine
| Autor: | Tsubasa Kato, Kazuaki Matsumura, Junya Ito, Maki Kamoshita, Suong-Hyu Hyon, Katsuyoshi Fujiwara, Naomi Kashiwazaki, Takafumi Namiki |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Embryology Swine lcsh:Medicine Cryopreservation Embryo Culture Techniques chemistry.chemical_compound 0302 clinical medicine Cryoprotective Agents Animal Cells Pregnancy Vitrification Polylysine lcsh:Science Mammals 030219 obstetrics & reproductive medicine Multidisciplinary Mammalian Genomics Chemistry Pregnancy Outcome Embryo Genomics medicine.anatomical_structure OVA embryonic structures Vertebrates Physical Sciences Female Stem cell Cellular Types Research Article Animal Types Materials Science Material Properties Embryonic Development Permeability Andrology 03 medical and health sciences medicine Genetics Animals Domestic Animals Blastocyst lcsh:R Embryogenesis Embryos Organisms Biology and Life Sciences Glutathione Cell Biology Embryo Transfer Sperm 030104 developmental biology Germ Cells Animal Genomics Amniotes Oocytes lcsh:Q Blastocysts Zoology Developmental Biology |
| Zdroj: | PLoS ONE PLoS ONE, Vol 12, Iss 4, p e0176711 (2017) |
| ISSN: | 1932-6203 |
| Popis: | Vitrification is a powerful tool for the efficient production of offspring derived from cryopreserved oocytes or embryos in mammalian species including domestic animals. Genome editing technologies such as transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated (Cas)9 are now available even for domestic species, suggesting that the vitrification of embryos at the pronuclear stage (PN) will be more important because they could provide genomic host cells to be targeted by TALENs or CRISPR/Cas9. Although we reported the successful production of piglets derived from vitrified PN embryos by a solid-surface vitrification method with glutathione supplementation, further improvements are required. The cryoprotective agent (CPA) carboxylated ε-poly-L-lysine (COOH-PLL) was introduced in 2009. COOH-PLL reduces the physical and physiological damage caused by cryopreservation in mammalian stem cells and the vitrification of mouse oocytes and embryos. Those results suggested that vitrification of COOH-PLL may help improve the developmental ability of pig embryos vitrified at the PN stage. However, it remains unclear whether COOH-PLL is available as a CPA for the vitrification of embryos in domestic species. In this study, we evaluated COOH-PLL as a CPA with ethylene glycol (EG) and Cryotop as a device for the vitrification of PN pig embryos. Exposure to vitrification solution supplemented with COOH-PLL up to 30% did not decrease developmental ability to the 2-cell stage and the blastocyst stage. After warming, most of the vitrified embryos survived regardless of the concentration of COOH-PLL (76.0 ± 11.8% to 91.8 ± 4.6%). However, the vitrified embryos without COOH-PLL showed a lower development rate up to the blastocyst stage (1.3 ± 1.0%) compared to the fresh embryos (28.4 ± 5.0%) (p |
| Databáze: | OpenAIRE |
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