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Department of Cell Biology,3 Harvard Medical School, Boston, Massachusetts 02115
Center for Engineering in Medicine and Surgical Services,4 Massachusetts General Hospital, Harvard Medical School and Shriners Hospital for Children, Boston, Massachusetts 02114
Transgenic Mouse Facility,5 Beth Israel Deaconess Medical Center, Boston, Massachusetts 02115
Department of Mechanical Engineering,6 University of Massachusetts, Dartmouth, Massachusetts 02747
Mouse sperm with and without trehalose were desiccated under nitrogen gas and stored at 4°C and 22°C. After rehydration, sperm were injected into oocytes using intracytoplasmic sperm injection and embryonic development was followed. Sperm were dried for 5.0, 6.25, or 7.5 min, stored at 22°C for 1 wk with and without trehalose. The percentages of blastocysts that developed from sperm with trehalose were 51%, 31%, and 20%, respectively, which was significantly higher than sperm without trehalose (10%, 3%, and 5%, respectively). Desiccation and storage in medium with trehalose significantly increased sperm developmental potential compared to medium without trehalose. Sperm dried for 5 min produced more blastocysts than sperm dried for 6.25 or 7.5 min. When sperm were dried in trehalose for 5 min and stored for 1 wk, 2 wk, 1 mo, or 3 mo at 4°C, the percentages of blastocysts were 73%, 84%, 63%, and 39%; whereas those stored at 22°C for 1 wk, 2 wk, or 1 mo were significantly lower (53%, 17%, and 6%, respectively). Embryos from sperm partially desiccated in trehalose for 5 min and stored at 4°C for 1 or 3 mo were transferred to 10 pseudopregnant recipients. Implantation rates were 81% and 48%; live fetuses were 26% and 5%, respectively. One of the recipients delivered three live fetuses. The results show that trehalose has a significant beneficial effect in preserving the developmental potential of mouse sperm following partial desiccation and storage at temperatures above freezing.
embryo, fertilization, gamete biology, ICSI, sperm, sperm desiccation, trehalose
2 Correspondence: John D. Biggers, Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115. FAX: 781 274 9988; john_biggers{at}hms.harvard.edu
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