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Desiccation Tolerance of Spermatozoa Dried at Ambient Temperature: Production of Fetal Mice¹

Center for Engineering in Medicine,³ Massachusetts General Hospital, Harvard Medical School, and Shriners Hospital for Children, Boston, Massachusetts 02115 Department of Cell Biology,⁴ Harvard Medical School, Boston, Massachusetts 02115 Beth Israel Deaconess Medical Center,⁵ Harvard Medical School, Boston, Massachusetts 02115

Long-term preservation of mouse sperm by desiccation is economically and logistically attractive. The current investigation is a feasibility study of the preservation of mouse sperm by convective drying in an inert gas (nitrogen). Mouse sperm from the B6D2F1 strain isolated in an EGTA-supplemented Tris-HCl buffer were dried using three different drying rates and were stored for 18–24 h at 4°C. The mean final moisture content was <5% for all the protocols. After intracytoplasmic sperm injection (ICSI), the mean blastocyst formation rates were 64%, 58%, and 35% using the rapid-, moderate-, and slow-drying protocols, respectively. The slow-drying protocol resulted in a rate of development significantly lower than that observed using rapid- and moderate-drying protocols and indicated that a slower drying rate may be detrimental to the DNA integrity of mouse sperm. The transfer of 85 two- or four-cell embryos that were produced using rapidly desiccated sperm resulted in 11 fetuses (13%) on Day 15 compared with the production of 34 fetuses (40%) produced using the transfer of 86 two- or four-cell embryos that were produced using fresh sperm (P < 0.05). The results demonstrate the feasibility of using a convective drying protocol for the successful desiccation of mouse sperm and identifies some of the important parameters required for optimization of the procedure.

¹ This work was supported by the NIH Cooperative Program on mouse sperm 5U01HD38227-03.

² The first two authors (S.B and L.Z.) contributed equally to this work

³ Correspondence: Mehmet Toner, Massachusetts General Hospital, SHC, HMS, 51 Blossom Street, Boston, MA 02114. FAX: 617 371 4950; mtoner{at}sbi.org

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