Ovariectomy and ovarian tissue cryopreservation has the potential to preserve the natural fertility of cancer patients prior to sterilizing chemo- and radiotherapies. Ovarian tissue cryopreservation with the conventional slow-freezing method has yielded limited success, partly because of oocyte loss during freeze-thaw and subsequent transplant. Based on the high-efficiency vitrification Cryotop method, a practical vitrification procedure for murine, bovine and human ovarian tissue was developed. A Cryotissue method was designed for cryopreservation of ovarian tissue, and vitrification experiments were performed in a bovine animal model with ovarian size and structure similar to the human. There was no difference in oocyte viability (>89%) between fresh and vitrified ovarian cortical tissue in either bovine or human samples. Ovarian tissue was successfully autotransplanted to six cattle. Autotransplantation of vitrified-warmed tissue back to the cattle resulted in no loss of oocyte viability. In addition, human ovarian tissue from cancer patients, and from ovary transplant donors was also vitrified by the Cryotissue method. After warming, high oocyte survival in human tissue (similar to bovine tissue) was obtained. These results indicate that an ultra-rapid cooling vitrification method has the potential for clinical use in human ovarian tissue cryopreservation.