{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T22:58:15Z","timestamp":1780613895128,"version":"3.54.1"},"reference-count":29,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2005,2,4]],"date-time":"2005-02-04T00:00:00Z","timestamp":1107475200000},"content-version":"vor","delay-in-days":4358,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecular Reproduction Devel"],"published-print":{"date-parts":[[1993,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Bovine blastocysts were produced through maturation, fertilization, and development in vitro. For vitrification, solutions designated EFS, GFS, and PFS were prepared; these were 40% ethylene glycol, 40% glycerol, and 40% propylene glycol, respectively, diluted in modified phosphate\u2010buffered saline (PBS) containing 30% Ficoll + 0.5 M sucrose. The embryos were exposed to the solutions in one step at room temperature, kept in the solutions for various times, vitrified in liquid nitrogen, and warmed rapidly. When the embryos were vitrified in EFS solution after 1 or 2 min exposure, the postwarming survival rate, assessed by the reexpansion of the blastocoel, was 74\u201377%. However, when the exposure time was extended to 3 min or longer, this rate dropped to 7\u20130%. This reduction was attributed to the toxicity of ethylene glycol. Of the embryos vitrified in GFS solution, 53% survived when they were cooled after 1 min exposure; as the duration of the exposure increased, the survival rate increased, reaching a peak (72%) at 4 min. The rate then decreased gradually with exposure time. In PFS solution, embryos surviving after vitrification were recovered only with 1 min exposure (33%), reflecting the high toxicity of propylene glycol. After vitrification in EFS or GFS solution, two embryos were nonsurgically transferred into each of 14 recipient animals. Of the 14 recipients, ten (71%) became pregnant; two resulted in early stillbirths, four recipients delivered twins (four alive and four stillborn), and two delivered single live calves, demonstrating the effectiveness of this simple vitrification method for the cryopreservation of in\u2010vitro\u2010produced bovine blastocysts. \u00a9 1993 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/mrd.1080340306","type":"journal-article","created":{"date-parts":[[2005,2,23]],"date-time":"2005-02-23T23:58:57Z","timestamp":1109203137000},"page":"266-271","source":"Crossref","is-referenced-by-count":93,"title":["Successful vitrification of bovine blastocysts, derived by in vitro maturation and fertilization"],"prefix":"10.1002","volume":"34","author":[{"given":"S.","family":"Tachikawa","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"T.","family":"Otoi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S.","family":"Kondo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"T.","family":"Machida","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.","family":"Kasai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2005,2,4]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod12.2.260"},{"key":"e_1_2_1_3_1","doi-asserted-by":"publisher","DOI":"10.1262\/jrd1977.36.69"},{"key":"e_1_2_1_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/0011-2240(84)90079-8"},{"key":"e_1_2_1_5_1","doi-asserted-by":"publisher","DOI":"10.1136\/vr.122.12.282"},{"key":"e_1_2_1_6_1","doi-asserted-by":"publisher","DOI":"10.1530\/jrf.0.0830753"},{"key":"e_1_2_1_7_1","doi-asserted-by":"publisher","DOI":"10.1530\/jrf.0.0900279"},{"key":"e_1_2_1_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/0093-691X(91)90193-H"},{"key":"e_1_2_1_9_1","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod46.6.1042"},{"key":"e_1_2_1_10_1","doi-asserted-by":"publisher","DOI":"10.1530\/jrf.0.0890091"},{"key":"e_1_2_1_11_1","doi-asserted-by":"publisher","DOI":"10.1530\/jrf.0.0630175"},{"key":"e_1_2_1_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/0093-691X(93)90326-Z"},{"key":"e_1_2_1_13_1","first-page":"165","article-title":"Cryopreservation of bovine blastocysts developed from in vitro fertilized oocytes matured in vitro","volume":"13","author":"Kuwayama M","year":"1991","journal-title":"Jpn J Anim Reprod Technol"},{"key":"e_1_2_1_14_1","first-page":"240","article-title":"Vitrification of mouse 8\u2010cell embryos with glycerol as a cryoprotectant","volume":"36","author":"Liehman P","year":"1990","journal-title":"Folia Biol"},{"key":"e_1_2_1_15_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1439-0531.1989.tb00447.x"},{"key":"e_1_2_1_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/0093-691X(87)90071-9"},{"key":"e_1_2_1_17_1","first-page":"270","article-title":"Pregnancies following transfer of cattle embryos preserved by vitrification","volume":"7","author":"Massip A","year":"1986","journal-title":"Cryo\u2010Letters"},{"key":"e_1_2_1_18_1","first-page":"23","article-title":"Survival of mouse embryos derived from in vitro fertilization after ultrarapid freezing and thawing","volume":"6","author":"Nakagata N","year":"1989","journal-title":"J Mammal Ova Res"},{"key":"e_1_2_1_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/0093-691X(91)90151-3"},{"key":"e_1_2_1_20_1","doi-asserted-by":"publisher","DOI":"10.1038\/313573a0"},{"key":"e_1_2_1_21_1","first-page":"260","article-title":"A simple and efficient procedure for preservation of mouse embryos by vitrification","volume":"7","author":"Scheffen B","year":"1986","journal-title":"Cryo\u2010Letters"},{"key":"e_1_2_1_22_1","doi-asserted-by":"publisher","DOI":"10.1071\/RD9910621"},{"key":"e_1_2_1_23_1","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod39.3.546"},{"key":"e_1_2_1_24_1","unstructured":"SuzukiT IshidaT SakaiY(1988):Twinning of ipsilateral and bilateral transfer of frozen bovine embryos. 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