title: Successful vitrification of bovine blastocysts on paper container.
abstract: Cryopreservation of bovine embryos can be performed by a variety of methods with variable degree of success. Here, we report a new, easy to perform, simple, inexpensive, and successful method for vitrification of bovine blastocysts. In vitro produced bovine blastocysts were exposed to vitrification solution (5.5 m ethylene glycol, 10% serum and 1% sucrose) in one single step for 20 s, loaded on a paper container prepared from commonly available non-slippery, absorbent writing paper, and then were directly plunged into liquid nitrogen for storage. Vitrified blastocysts were warmed by serial rinsing in 0.5, 0.25 and 0.125 m sucrose solution for 1 min each. Results showed that one step exposure of bovine blastocysts to cryoprotective agents was sufficient to achieve successful cryopreservation. Under these conditions, more than 95% of blastocysts survived the vitrification-warming on paper containers which was significantly higher than those obtained from other containers, such as electron microscope (EM) grid (78.1%), open pulled straw (OPS; 80.2%), cryoloop (76.2%) or plastic straw (73.9%). Embryo transfer of blastocysts vitrified-warmed on paper container resulted in successful conception (19.3%) and full-term live birth of offspring (12.3%) which were lower (P < 0.05) than those obtained from non-vitrified blastocysts (38.0 and 32.7%) but were comparable (P > 0.05) to those obtained from blastocysts vitrified-warmed on EM grid (23.3 and 14.2%). Our results, therefore, suggest that paper may be an inexpensive and useful container for the cryopreservation of animal embryos.
authors: Department of Animal Science and Biotechnology, Sangji Youngseo College, Wonju 220-713, South Korea.
authors: Y M
authors: YM
authors: Kim
authors: S J
authors: SJ
authors: Uhm
authors: M K
authors: MK
authors: Gupta
authors: J S
authors: JS
authors: Yang
authors: J-G
authors: JG
authors: Lim
authors: Z C
authors: ZC
authors: Das
authors: Y T
authors: YT
authors: Heo
authors: H-J
authors: HJ
authors: Chung
authors: I-K
authors: IK
authors: Kong
authors: N-H
authors: NH
authors: Kim
authors: H T
authors: HT
authors: Lee
authors: D H
authors: DH
authors: Ko
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D002417:Cattle / Q000196:embryology* / Q000502:physiology; D015925:Cryopreservation / Q000662:veterinary*; D046149:Embryo Culture Techniques; D005260:Female; D005307:Fertilization in Vitro; D059471:In Vitro Oocyte Maturation Techniques; D010209:Paper; D011247:Pregnancy