SignificanceIatrogenic failures of assisted reproduction technology could be associated with routine sperm preparation techniques. Limitations of conventional sperm selection methods include the inability to efficiently sort functional spermatozoa and assess sperm fertilization potential. We developed a robust microfluidic sperm sorting system by using a diffuser-type microfluidic sperm sorter device capable of ultrahigh-throughput selection and separation of motile, DNA-intact, and functionally competent sperm. The strategy inclusively targeted the intrinsic traits related to fertility and successfully produced livebirths from low-dose insemination of microfluidic sorted spermatozoa. The fertile subpopulation was identified based on the kinetic and trajectory patterns as the sinuous, transitional cohort. The clinical significance of microfluidic sperm sorting is reflected by the established pregnancy and live births of calves.

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A membrane-bound inositol phospholipid-specific phospholipase C was solubilized from rice (Oryza sativa L.) microsomal membranes and purified to apparent homogeneity using a series of chromatographic separations. l h e apparent molecular mass of the enzyme was estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 42,000 D, and the isoelectric point was 5

Abstract. The viability of reconstituted embryos derived from bovine somatic cells were evaluated. A total of 54 fetal somatic cell-derived embryos, in which some embryos were previously cryopreserved, were transferred to 12 recipient cattle (total 26 times). Three of 12 cattle that received fresh embryos became pregnant and delivered 3 calves (1 aborted at 223d and 2 live born), whereas none out of the 5 recipients became pregnant after transfer of embryos cryopreserved by slow cooling. Of 9 cattle that received embryos vitrificated 2 became pregnant. One of these two vitrification-derived embryos died during early pregnancy and another developed to term but was stillborn. These aborted fetuses, stillborn calf and two live born calves were identified as being derived from donor cells. The growth of live born calves showed no abnormalities when examined at 8 months of age. It was therefore concluded that: reconstituted embryos derived from fibroblast cells from female fetus could develop to term, that vitrification might be available to preserve these embryos and that transfer of these fresh embryos resulted in viable offspring with normal growth.

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