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This Article Full Text Full Text (PDF) All Versions of this Article: 68/3/881    most recent biolreprod.102.007948v4 biolreprod.102.007948v3 biolreprod.102.007948v2 biolreprod.102.007948v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal My Folders Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Migishima, F. Articles by Yokoyama, M. Search for Related Content PubMed PubMed Citation Articles by Migishima, F. Articles by Yokoyama, M. Agricola Articles by Migishima, F. Articles by Yokoyama, M.

Successful Cryopreservation of Mouse Ovaries by Vitrification¹

^a Mitsubishi Kagaku Institute of Life Sciences, Minamiooya 11, Machida, Tokyo 194-8511, Japan ^b Department of Obstetrics and Gynecology, Kitasato University School of Medicine, Sagamihara, Kanagawa 228-8555, Japan

We developed a new method of cryopreservation of whole ovaries by vitrification using DAP213 (2 M dimethyl sulfoxide, 1 M acetamide, and M propylene glycol) as a cryoprotectant. Four-week-old C57BL/6 mice that underwent partial ovariectomy were orthotopically transplanted with cryopreserved or fresh ovaries (experimental or control group) isolated from 10-day-old green fluorescent protein (GFP)-transgenic mice (+/+). GFP-positive pups were similarly obtained from both groups by natural mating or in vitro fertilization (IVF) followed by embryo transfer, indicating that the cryopreserved ovaries by vitrification retain their fecundity. However, a statistically significant difference (P < 0.05) was found between both groups with respect to the following parameters: the number of GFP-positive pups born by natural mating/grafted ovary (0.8 ± 0.3 for the experimental group versus 2.0 ± 0.7 for the control group, mean ± SEM), the number of collected oocytes by superovulation per mouse (7.0 ± 1.7 for the experimental group versus 22.7 ± 3.2 for the control group), the percentage of two-cell embryos obtained from GFP-positive oocytes by IVF (38.5% for the experimental group versus 90.0% for the control group). Histologically, normal development of follicles and formation of corpora lutea were observed in frozen-thawed grafts. However, estimated number of follicles decreased in frozen-thawed ovaries compared with fresh ovaries. Taken together, cryopreservation of the ovary by vitrification seems a promising method to preserve ovarian function, but further studies are required to overcome the possible inhibitory effects of this method on the growth of the ovarian graft.

¹ This study was partially supported by Special Coordination Funds of the Japanese Ministry of Education, Culture, Sports, Science, and Technology.

² Correspondence: Minesuke Yokoyama, Mitsubishi Kagaku Institute of Life Sciences, Minamiooya 11, Machida, Tokyo 194-8511, Japan. FAX: 81 42 724 6316; e-mail: myoko{at}libra.ls.m-kagaku.co.jp

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