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1. Successful vitrification of bovine blastocysts on paper container

Author

Hoon Taek Lee, Young-Tae Heo, S.J. Uhm, Y.M. Kim, N.-H. Kim, Hak-Jae Chung, Il-Keun Kong, D.H. Ko, J.-G. Lim, J.S. Yang, Mukesh Kumar Gupta, and Ziban Chandra Das

Subjects
Paper, Single step, Fertilization in Vitro, Cryopreservation, Embryo Culture Techniques, Andrology, chemistry.chemical_compound, Food Animals, Embryo cryopreservation, Pregnancy, Animals, Vitrification, Bovine embryo, Small Animals, Equine, Chemistry, business.industry, Straw, Embryo transfer, In Vitro Oocyte Maturation Techniques, Biotechnology, Blastocyst, embryonic structures, Cattle, Female, Animal Science and Zoology, business, Ethylene glycol
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 (P0.05) than those obtained from non-vitrified blastocysts (38.0 and 32.7%) but were comparable (P0.05) 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.

Published
2012

2. Successful vitrification of bovine blastocysts on paper container.

Author

Kim YM, Uhm SJ, Gupta MK, Yang JS, Lim JG, Das ZC, Heo YT, Chung HJ, Kong IK, Kim NH, Lee HT, and Ko DH

Subjects
Animals, Cattle physiology, Embryo Culture Techniques, Female, Fertilization in Vitro, In Vitro Oocyte Maturation Techniques, Paper, Pregnancy, Blastocyst physiology, Cattle embryology, Cryopreservation veterinary
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., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Published
2012
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