Mouse Sperm Desiccated and Stored in Trehalose Medium Without Freezing1

Abstract

Mouse sperm with and without trehalose were desiccated under nitrogen gas and stored at 4 degrees C and 22 degrees 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 degrees 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 degrees C, the percentages of blastocysts were 73%, 84%, 63%, and 39%; whereas those stored at 22 degrees 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 degrees 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.

Effects of freeze-drying on cytology, ultrastructure, DNA fragmentation, and fertilizing ability of bovine sperm

“…albumin [7] and [12], EGTA solution [13] and [14] and trehalose [15]. However, it is not clear how freeze-drying affects specific components of sperm cells.…”

The Effect of Freeze Drying Using Different Media and Storage Temperatures on Some Parameters of Buffalo Bull Spermatozoa

“…The sperm plasma membrane and acrosome integrities are highly susceptible to injuries, due to loss of water during dehydration (Sherman, 1954, Yushchenko, 1957, Kusakabe et al, 2001, McGinnis et al, 2005and Kawase & Suzuki, 2011. Water loss from phospholipid head groups in cell membranes could be due to lateral phase separation resulting in extravasation of intracellular contents (Crowe et al, 1997).…”

“…Many trials were carried out to protect sperm structures during freeze-drying via various protecting substances, e.g. albumin , EGTA (Kusakabe et al, 2001 andKaneko et al, 2003), EDTA (Kaneko and Serikawa, 2012) and trehalose (McGinnis et al, 2005). The main target of the current study was to investigate the effect of freeze-drying using different media and storage temperatures on some parameters of buffalo bull spermatozoa.…”

A catalyst for change in reproductive science: John D. Biggers as a mentor’s mentor

“…The result was amazing. Sperm dried to 0.3 % water can be stored at 4C for months, rehydrated with water, injected into eggs and produce live offspring [31][32][33][34][35][36].…”