Hollow Fiber Vitrification: A Novel Method for Vitrifying Multiple Embryos in a Single Device
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概要
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Current embryo vitrification methods with proven efficacy are based on the minimum volume cooling (MVC) concept by which embryos are vitrified and rewarmed ultrarapidly in a very small amount of cryopreserving solution to ensure the high viability of the embryos. However, these methods are not suitable for simultaneously vitrifying a large number of embryos. Here, we describe a novel vitrification method based on use of a hollow fiber device, which can easily hold as many as 40 mouse or 20 porcine embryos in less than 0.1 μl of solution. Survival rates of up to 100% were obtained for mouse embryos vitrified in the presence of 15% DMSO, 15% ethylene glycol and 0.5 M sucrose using the hollow fiber vitrification (HFV) method, regardless of the developmental stage of the embryos (1-cell, 2-cell, morula or blastocyst; n = 50/group). The HFV method was also proven to be effective for vitrifying porcine in vitro- and in vivo-derived embryos that are known to be highly cryosensitive. For porcine embryos, the blastocyst formation rate of in vitro maturation (IVM)-derived parthenogenetic morulae after vitrification (48/65, 73.8%) did not decrease significantly compared with non-vitrified embryos (59/65, 90.8%). Transfer of 72 in vivo-derived embryos vitrified at the morula/early blastocyst stages to 3 recipients gave rise to 29 (40.3%) piglets. These data demonstrate that the HFV method enables simultaneous vitrification of multiple embryos while still adhering to the MVC concept, and this new method is very effective for cryopreserving embryos of mice and pigs.
著者
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Ikezawa Yuka
Laboratory Of Developmental Engineering Department Of Life Sciences School Of Agriculture Meiji Univ
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SASAYAMA Norihisa
Research & Development Institute for Artificial Organs, Research & Development Laboratory, Nipro Cor
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SHIRASU Akio
Research & Development Institute for Artificial Organs, Research & Development Laboratory, Nipro Cor
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Matsunari Hitomi
Laboratory Of Developmental Engineering Department Of Life Sciences School Of Agriculture Meiji Univ
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Nagashima Hiroshi
Laboratory For Evolutionary Morphology Riken Cdb
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Nakano Kazuaki
Laboratory Of Developmental Engineering Department Of Life Sciences School Of Agriculture Meiji Univ
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Takahashi Masashi
Department Of Animal And Grassland Research National Agricultural Center For Kyushu Okinawa Region
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MAEHARA Miki
Laboratory of Developmental Engineering, Department of Life Sciences, School of Agriculture, Meiji University
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HAGIWARA Yui
Research & Development Institute for Artificial Organs, Research & Development Laboratory, Nipro Corporation
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OHTA Hisayoshi
Tokyo Development Foundation for Agriculture, Forestry and Fisheries
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HAGIWARA Yui
Research & Development Institute for Artificial Organs, Research & Development Laboratory, Nipro Corporation, Shiga 525-0055, Japan
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MAEHARA Miki
Laboratory of Developmental Engineering, Department of Life Sciences, School of Agriculture, Meiji University, Kawasaki 214-8571, Japan
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SASAYAMA Norihisa
Research & Development Institute for Artificial Organs, Research & Development Laboratory, Nipro Corporation, Shiga 525-0055, Japan
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SHIRASU Akio
Research & Development Institute for Artificial Organs, Research & Development Laboratory, Nipro Corporation, Shiga 525-0055, Japan
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