Wild-derived mice have long offered invaluable experimental models for mouse genetics because of their high evolutionary divergence from laboratory mice. A number of wild-derived strains are available from the RIKEN BioResource Center (BRC), but they have been maintained as living stocks because of the unavailability of assisted reproductive technology (ART). In this study, we sought to devise ART for 37 wild-derived strains from five subspecies of Mus musculus maintained at the BRC. Superovulation of females was effective (more than 15 oocytes per female) for 34 out of 37 strains by treatment with either equine chorionic gonadotropin or anti-inhibin serum, depending on their genetic background (subspecies). The collected oocytes could be fertilized in vitro at mean rates of 79.0% and 54.6% by the optimized protocol using fresh or frozen-thawed spermatozoa, respectively. They were cryopreserved at the 2-cell stage by vitrification with an ethylene glycol-based solution. In total, 94.6% of cryopreserved embryos survived the vitrification procedure and restored their normal morphology after warming. A conventional embryo transfer protocol could be applied to 25 out of the 35 strains tested. In the remaining 10 strains, live offspring could be obtained by a modified embryo transfer protocol using cyclosporin A treatment and co-transfer of ICR (laboratory mouse strain) embryos. Thus, ART for 37 wild-derived strains was devised successfully and is now routinely used for their preservation and transportation. The information provided here might facilitate broader use and wider distribution of wild-derived mice for biomedical research.
Diamond-like amorphous carbon films doped with nitrogen (DAC:N) were deposited on Si and glass wafers intermittently using i-C4H10/N2 repetitive supermagnetron plasma chemical vapor deposition. Deposition duration, which is equal to a plasma heating time of wafer, was selected to be 40 or 60 s, and several layers were deposited repetitively to form one thick film. DAC:N films were deposited at a lower-electrode temperature of 100 °C as a function of upper- and lower-electrode rf powers (200 W/200 W–1 kW/1 kW) and N2 concentration (0%–80%). With an increase in N2 concentration and rf power, the resistivity and the optical band gap decreased monotonously. With increase of the deposition duration from 40 to 60 s, resistivity decreased to 0.03Ω cm and optical band gap decreased to 0.02 eV (substantially equal to 0 eV within the range of experimental error), at an N2 concentration of 80% and rf power of 1 kW(/1 kW).
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