Cryoloop vitrification yields superior survival of Rhesus monkey blastocysts

R. R. Yeoman, B. Gerami-Naini, S. Mitalipov, K. D. Nusser, A. A. Widmann-Browning, D. P. Wolf

    Research output: Contribution to journalArticlepeer-review

    50 Scopus citations


    Background: Vitrification using the cryoloop procedure was evaluated for preservation of non-human primate blastocysts by comparing survival results from two different cryoprotectant mixtures with prior results from controlled rate cooling. Methods: Rhesus monkey blastocysts were produced by intracytoplasmic sperm injection of mature oocytes from cycling females stimulated with recombinant human hormones. Morphologically well-formed blastocysts were divided between Procedure A (2.8 mol/l dimethylsulphoxide and 3.6 mol/l ethylene glycol with 0.65 mol/l sucrose and 25 μmol/l Ficoll in TALP-HEPES with 20% fetal bovine serum (TH20)) and Procedure B (3.4 mol/l glycerol and 4.5 mol/l ethylene glycol in TH20). After >48 h in liquid nitrogen, the removal of cryoprotectants was accomplished in the presence of a 3-step series of decreasing sucrose concentrations in TH20. Surviving embryos were co-cultured on buffalo rat liver cells. Results: Of 16 blastocysts vitrified via Procedure A, 38% survived with minimal lysis and only one hatched in culture; in contrast, of 33 blastocysts vitrified by Procedure B, 85% survived and 71% hatched. Of 22 blastocysts cryopreserved by conventional slow cooling, 36% survived and 6% hatched. Transfer into three recipients, each with two embryos vitrified with Procedure B, resulted in a successful twin-term pregnancy. Conclusion: Modified cryoloop vitrification with a final solution of 3.4 mol/l glycerol and 4.5 mol/l ethylene glycol is a promising procedure for preserving Rhesus monkey blastocysts that is simple, rapid, and inexpensive.

    Original languageEnglish (US)
    Pages (from-to)1965-1969
    Number of pages5
    JournalHuman Reproduction
    Issue number9
    StatePublished - 2001


    • Blastocysts
    • Cryoloop
    • Cryopreservation
    • Monkey
    • Vitrification

    ASJC Scopus subject areas

    • Reproductive Medicine
    • Obstetrics and Gynecology


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