Comparison of the efficacy of conventional slow freezing and rapid cryopreservation methods for bovine embryos

A.M. van Wagtendonk-de Leeuw, J.H. den Daas, T.A. Kruip, W.F. Rail

    Research output: Contribution to journalArticleAcademicpeer-review

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    Abstract

    Day 7 bovine morulae and early blastocysts were randomly assigned to one of four cryopreservation methods: (i) a modified conventional controlled slow freezing and stepwise dilution after thawing; and three methods which enable direct transfer of the embryo into the recipient upon thawing: (ii) conventional controlled slow freezing and a modification of a one-step procedure, (iii) vitrification with 6.5 M glycerol plus 6% BSA (w/v), and (iv) vitrification with 25% glycerol (v/v) and 25% propanediol (v/v). In a comparative in vitro study, the percentage of grade 1 and 2 embryos developing into expanded blastocysts in culture for cryopreservation methods 1-4 were, respectively, 53% (29/55), 33% (20/61), 44% (26/59), and 51% (17/33). Method 2 yielded a significantly lower survival rate than methods 1 (P < 0.1) and 4 (P < 0.05) and was excluded from a subsequent test of in vivo development. Pregnancy rates (Day 60) after transfer of embryos cryopreserved by methods 1,3, and 4 were, respectively, 59% (20/34), 43% (17/40), and 24% (5/21). Method 4 yielded a significantly lower pregnancy rate than method 1 (P < 0.05). Method 3, however, did not yield a statistically different pregnancy rate (P > 0.1) when compared to method 1. Method 3 has considerable promise in providing a successful method for the cryopreservation of bovine embryos that (i) reduces the time required for equilibration and cooling, (ii) provides for simple and rapid one-step dilution of cryoprotectant after thawing, and (iii) enables more embryos to be thawed and transferred per unit time.
    Original languageEnglish
    Pages (from-to)157-167
    JournalCryobiology
    Volume32
    Issue number2
    DOIs
    Publication statusPublished - 1995

    Fingerprint

    Thawing
    Cryopreservation
    Freezing
    cryopreservation
    Vitrification
    embryo (animal)
    freezing
    Embryonic Structures
    Glycerol
    Dilution
    cattle
    Propylene Glycols
    thawing
    vitrification
    methodology
    blastocyst
    Blastocyst
    Cooling
    glycerol
    propanediols

    Cite this

    van Wagtendonk-de Leeuw, A.M. ; den Daas, J.H. ; Kruip, T.A. ; Rail, W.F. / Comparison of the efficacy of conventional slow freezing and rapid cryopreservation methods for bovine embryos. In: Cryobiology. 1995 ; Vol. 32, No. 2. pp. 157-167.
    @article{b556f1d49cbd4ba7b1198b140f99ac36,
    title = "Comparison of the efficacy of conventional slow freezing and rapid cryopreservation methods for bovine embryos",
    abstract = "Day 7 bovine morulae and early blastocysts were randomly assigned to one of four cryopreservation methods: (i) a modified conventional controlled slow freezing and stepwise dilution after thawing; and three methods which enable direct transfer of the embryo into the recipient upon thawing: (ii) conventional controlled slow freezing and a modification of a one-step procedure, (iii) vitrification with 6.5 M glycerol plus 6{\%} BSA (w/v), and (iv) vitrification with 25{\%} glycerol (v/v) and 25{\%} propanediol (v/v). In a comparative in vitro study, the percentage of grade 1 and 2 embryos developing into expanded blastocysts in culture for cryopreservation methods 1-4 were, respectively, 53{\%} (29/55), 33{\%} (20/61), 44{\%} (26/59), and 51{\%} (17/33). Method 2 yielded a significantly lower survival rate than methods 1 (P < 0.1) and 4 (P < 0.05) and was excluded from a subsequent test of in vivo development. Pregnancy rates (Day 60) after transfer of embryos cryopreserved by methods 1,3, and 4 were, respectively, 59{\%} (20/34), 43{\%} (17/40), and 24{\%} (5/21). Method 4 yielded a significantly lower pregnancy rate than method 1 (P < 0.05). Method 3, however, did not yield a statistically different pregnancy rate (P > 0.1) when compared to method 1. Method 3 has considerable promise in providing a successful method for the cryopreservation of bovine embryos that (i) reduces the time required for equilibration and cooling, (ii) provides for simple and rapid one-step dilution of cryoprotectant after thawing, and (iii) enables more embryos to be thawed and transferred per unit time.",
    author = "{van Wagtendonk-de Leeuw}, A.M. and {den Daas}, J.H. and T.A. Kruip and W.F. Rail",
    year = "1995",
    doi = "10.1006/cryo.1995.1014",
    language = "English",
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    Comparison of the efficacy of conventional slow freezing and rapid cryopreservation methods for bovine embryos. / van Wagtendonk-de Leeuw, A.M.; den Daas, J.H.; Kruip, T.A.; Rail, W.F.

    In: Cryobiology, Vol. 32, No. 2, 1995, p. 157-167.

    Research output: Contribution to journalArticleAcademicpeer-review

    TY - JOUR

    T1 - Comparison of the efficacy of conventional slow freezing and rapid cryopreservation methods for bovine embryos

    AU - van Wagtendonk-de Leeuw, A.M.

    AU - den Daas, J.H.

    AU - Kruip, T.A.

    AU - Rail, W.F.

    PY - 1995

    Y1 - 1995

    N2 - Day 7 bovine morulae and early blastocysts were randomly assigned to one of four cryopreservation methods: (i) a modified conventional controlled slow freezing and stepwise dilution after thawing; and three methods which enable direct transfer of the embryo into the recipient upon thawing: (ii) conventional controlled slow freezing and a modification of a one-step procedure, (iii) vitrification with 6.5 M glycerol plus 6% BSA (w/v), and (iv) vitrification with 25% glycerol (v/v) and 25% propanediol (v/v). In a comparative in vitro study, the percentage of grade 1 and 2 embryos developing into expanded blastocysts in culture for cryopreservation methods 1-4 were, respectively, 53% (29/55), 33% (20/61), 44% (26/59), and 51% (17/33). Method 2 yielded a significantly lower survival rate than methods 1 (P < 0.1) and 4 (P < 0.05) and was excluded from a subsequent test of in vivo development. Pregnancy rates (Day 60) after transfer of embryos cryopreserved by methods 1,3, and 4 were, respectively, 59% (20/34), 43% (17/40), and 24% (5/21). Method 4 yielded a significantly lower pregnancy rate than method 1 (P < 0.05). Method 3, however, did not yield a statistically different pregnancy rate (P > 0.1) when compared to method 1. Method 3 has considerable promise in providing a successful method for the cryopreservation of bovine embryos that (i) reduces the time required for equilibration and cooling, (ii) provides for simple and rapid one-step dilution of cryoprotectant after thawing, and (iii) enables more embryos to be thawed and transferred per unit time.

    AB - Day 7 bovine morulae and early blastocysts were randomly assigned to one of four cryopreservation methods: (i) a modified conventional controlled slow freezing and stepwise dilution after thawing; and three methods which enable direct transfer of the embryo into the recipient upon thawing: (ii) conventional controlled slow freezing and a modification of a one-step procedure, (iii) vitrification with 6.5 M glycerol plus 6% BSA (w/v), and (iv) vitrification with 25% glycerol (v/v) and 25% propanediol (v/v). In a comparative in vitro study, the percentage of grade 1 and 2 embryos developing into expanded blastocysts in culture for cryopreservation methods 1-4 were, respectively, 53% (29/55), 33% (20/61), 44% (26/59), and 51% (17/33). Method 2 yielded a significantly lower survival rate than methods 1 (P < 0.1) and 4 (P < 0.05) and was excluded from a subsequent test of in vivo development. Pregnancy rates (Day 60) after transfer of embryos cryopreserved by methods 1,3, and 4 were, respectively, 59% (20/34), 43% (17/40), and 24% (5/21). Method 4 yielded a significantly lower pregnancy rate than method 1 (P < 0.05). Method 3, however, did not yield a statistically different pregnancy rate (P > 0.1) when compared to method 1. Method 3 has considerable promise in providing a successful method for the cryopreservation of bovine embryos that (i) reduces the time required for equilibration and cooling, (ii) provides for simple and rapid one-step dilution of cryoprotectant after thawing, and (iii) enables more embryos to be thawed and transferred per unit time.

    U2 - 10.1006/cryo.1995.1014

    DO - 10.1006/cryo.1995.1014

    M3 - Article

    VL - 32

    SP - 157

    EP - 167

    JO - Cryobiology

    JF - Cryobiology

    SN - 0011-2240

    IS - 2

    ER -