Preservation of murine embryos in a state of dormancy at 4°C

With the aim of improving preservation of blood products and organs for transplantation, we designed solutions to induce a state of dormancy in cells and tissues at 4 degreesC. The solutions were devoid of combinations of ions (e.g., K+, Rb+, Cs+, and NH+4 with HCO-3, H2PO-4, and Cl-) that are believed to break down low-density water in the entrance compartments of ion channels, resulting in cyclical open states (normal water) and closed states (low-density water). The total osmolality was always 0.29-0.3 osmol/kgH2O, made up of combinations of a di- or trisaccharide, a compatible solute, sodium sulfate, citrate, or chloride, and 1.75 mM CaCl2. The end point was the ability of murine embryos to progress to hatching in culture after preservation in such a solution at 4 degreesC. Embryos hatched after 5 or 6 days in some preservative solutions compared with 1-3 days in most saline solutions; survival was improved by pretreatment with sodium butyrate.