Transplantation of frozen-thawed mouse primordial follicles

Primordial follicles were isolated from juvenile mouse ovaries and cryopreserved by slow freezing with dimethylsulphoxide as the cryoprotectant. After thawing, approximately 80% of the oocytes and 65% of the somatic cells excluded Trypan Blue dye, indicating that cell membranes were still intact. Frozen-thawed cells were suspended in plasma clots and transplanted to the ovarian bursas of host animals that had been sterilized by oophorectomy. The grafts of frozen-thawed cells reorganized into morphologically distinguishable ovaries which produced signs of oestrogenic activity. After natural mating, host females produced normal offspring that were demonstrated by genetic markers to be derived from the transplanted frozen-thawed primordial follicles.