One limitation to the widespread use of in vitro–produced embryos in cattle is their poor survival following cryopreservation. Two approaches for enhancing survival of in vitro–produced bovine embryos following cryopreservation were evaluated: culture in the presence of hyaluronic acid and alterations in the cytoskeleton through cytochalasin B treatment. The experiment was a 2 × 2 factorial design to test main effects of hyaluronic acid added to culture at day 5 after insemination ( or −) and cryopreservation treatment (control or cytochalasin B). Embryos used for cryopreservation were blastocysts and expanded blastocysts harvested on day 7 after insemination. Cytochalasin B increased the percent of embryos that re-expanded (P < 0.0001) and that hatched following thawing (P < 0.05). The hatching percent was 29.6% for embryos treated with cytochalasin B versus 9.1% for control embryos. There was no significant effect of hyaluronic acid on survival although there was a tendency for embryos cultured with hyaluronic acid to have higher percent hatching if not treated with cytochalasin B (12.7% for hyaluronic acid versus 4.5% for control; hyaluronic acid × cytochalasin B interaction; P = 0.09). In conclusion, cytochalasin B treatment before freezing improved cryosurvival of bovine embryos produced in vitro. Such a treatment could be incorporated into methods for cryopreservation of bovine embryos provided post-transfer survival is adequate. In contrast, culture with hyaluronic acid was of minimal benefit— the increased cryosurvival in the absence of cytochalasin B was not sufficient to allow an adequate number of embryos to survive.