title: Development of a modified straw method for vitrification of in vitro-produced bovine blastocysts and various genes expression in between the methods.
abstract: This study evaluated a modified plastic straw loading method for vitrification of in vitro-produced bovine blastocysts. A modified straw was used with a depressed area on its inner surface to which embryos attach. In vitro-produced blastocysts were randomly assigned into three groups: (i) blastocysts attached to the inner surface of a plastic straw (aV), (ii) blastocysts attached to the inner surface of a modified plastic straw (maV), and (iii) non-vitrified blastocysts (control). The recovery rates were not significantly different between aV and maV groups (95.8% vs. 94.3%). The post-thaw survival rate did not significantly differ between aV and maV groups (86.4% vs. 88.2%). The total cell numbers of blastocyst was higher in control than in aV and maV groups (142 ± 21.8 vs. 117 ± 29.7 and 120 ± 25.2; P < 0.05), but not significantly differ between aV and maV groups. The mRNA levels of pro-apoptosis related genes Bax and Caspase-3 were higher in aV and maV than in control (P < 0.05). By contrast, the mRNA levels of anti-apoptotic genes Bcl-2 and Mcl-1 and of antioxidant-related genes MnSOD and Prdx5 were lower in aV and maV than in control (P < 0.05). Confocal microscopy analysis of Golgi apparatus and mitochondria showed that the fluorescence intensity of Golgi apparatus and mitochondria was higher in control than in aV and maV groups. In conclusion, both aV and maV methods can be used to successfully vitrify IVP blastocysts, with maV method to be preferable because of its easiness in embryo loading.
authors.affiliation: Department of Animal Science, Division of Applied Life Science, Gyeongsang National University, Jinju, Gyeongnam Province 660-701, Republic of Korea.
authors.forename: A-Na
authors.initials: AN
authors.lastname: Ha
authors.affiliation: Animal, Dairy, and Veterinary Sciences Department, Utah State University, Logan, UT 84322-4700, USA.
authors.forename: Sang-Ryeul
authors.initials: SR
authors.lastname: Lee
authors.affiliation: Animal, Dairy, and Veterinary Sciences Department, Utah State University, Logan, UT 84322-4700, USA.
authors.forename: Jeong-Seon
authors.initials: JS
authors.lastname: Jeon
authors.affiliation: Department of Animal Science, Division of Applied Life Science, Gyeongsang National University, Jinju, Gyeongnam Province 660-701, Republic of Korea.
authors.forename: Han-Seul
authors.initials: HS
authors.lastname: Park
authors.affiliation: Department of Animal Science, Division of Applied Life Science, Gyeongsang National University, Jinju, Gyeongnam Province 660-701, Republic of Korea.
authors.forename: Sang-Ho
authors.initials: SH
authors.lastname: Lee
authors.affiliation: Department of Animal Science, Division of Applied Life Science, Gyeongsang National University, Jinju, Gyeongnam Province 660-701, Republic of Korea.
authors.forename: Jong-In
authors.initials: JI
authors.lastname: Jin
authors.affiliation: Animal, Dairy, and Veterinary Sciences Department, Utah State University, Logan, UT 84322-4700, USA.
authors.forename: Benjamin R
authors.initials: BR
authors.lastname: Sessions
authors.affiliation: Animal, Dairy, and Veterinary Sciences Department, Utah State University, Logan, UT 84322-4700, USA.
authors.forename: Zhongde
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Animal, Dairy, and Veterinary Sciences Department, Utah State University, Logan, UT 84322-4700, USA; School of Veterinary Medicine, Utah State University, Logan, UT 84322-4700, USA; Center for Integrated Biosystems, Utah State University, Logan, UT 84322-4700, USA. Electronic address: ken.white@usu.edu.
authors.forename: Kenneth L
authors.initials: KL
authors.lastname: White
authors.affiliation: Department of Animal Science, Division of Applied Life Science, Gyeongsang National University, Jinju, Gyeongnam Province 660-701, Republic of Korea; Institute of Agriculture and Life Science, Gyeongsang National University, Jinju, Gyeongnam Province 660-701, Republic of Korea. Electronic address: ikong7900@gmail.com.
authors.forename: Il-Keun
authors.initials: IK
authors.lastname: Kong
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism*; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002417:Cattle; D004624:Embryo Transfer; D005260:Female; D005307:Fertilization in Vitro; D015870:Gene Expression; D006056:Golgi Apparatus / Q000378:metabolism; D008297:Male; D008928:Mitochondria / Q000378:metabolism; D064549:Myeloid Cell Leukemia Sequence 1 Protein / Q000235:genetics / Q000378:metabolism; D054464:Peroxiredoxins / Q000235:genetics / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D058989:Vitrification; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D064549:Myeloid Cell Leukemia Sequence 1 Protein; D051028:bcl-2-Associated X Protein; D054464:Peroxiredoxins; D013482:Superoxide Dismutase; D053148:Caspase 3
keywords: Bovine; IVP blastocysts; Modified plastic straw loading method; Post-thaw viability; Vitrification