M. Goldstein

Migration of fresh and cryopreserved human spermatozoa in polyacrylamide gel

The ability of freshly collected and frozen human spermatozoa to migrate in round capillary tubes containing specially formulated polyacrylamide gel was investigated, using 33 ejaculates from 27 donors. Each semen sample was divided; one portion was left undiluted, and the other portion was diluted to 50 x 10(6) sperm/ml. Glycerol was used as the cryoprotectant. The percentage of motile sperm cells was determined before and after freezing. Fresh semen contained a higher percentage of motile cells, which migrated farther than those of cryopreserved-thawed semen. Various correlations between the percentage of motile sperm and migration distance ranged from 0.57 to 0.62. There was a low positive correlation of migration distance with sperm cell concentration per milliliter, r = 0.25 to 0.34; and thus adjusting semen samples to a standard sperm concentration improved the accuracy of the test only slightly. The regression coefficient of migration distance on the percentage of motile sperm in fresh semen was 0.65, indicating that for each 10% increase in sperm motility, migration distance is predicted to increase 6.5 mm. Five batches of polyacrylamide gel gave uniform results, and the application of this stable gel to fertility investigations is discussed.

Sperm DNA fragmentation as treatment guidance for infertile couples

ported only in 2009. Isolation of these cells was replicated by a small number of other groups in mouse, rat and extended to a human model. The objective of this study is to demonstrate the presence of OSC in non-human primates (NHPs), which can generate mature oocytes following transplantation into the ovary. DESIGN: Eggs from ovarian stem cell transplantation were characterized and explored their fertilization potential. MATERIALS AND METHODS: The ovarian cortex was digested with collagenase IV and DNase followed by FACS with the DDX4 antibody. The cells were expanded in culture, transfected with a GFP lentivirus, and transplanted into the remaining ovary of the rhesus monkey. Following transplantation, gonadotropins were used for ovarian hyperstimulation to isolate oocytes. Oocytes transplant derivations were assessed by fluorescence, PCR and nested PCR. True oocyte phenotype with appearance of intact zona pellucida and polar body was observed along with expression of oocyte specific genes. Intracytoplasmic sperm injection was performed with collectedMII oocytes and fertilization and embryo development potential assessed. RESULTS: Here we demonstrate, for the first time, the presence of adult stem cells in the primate ovary which form mature oocytes with fertilization capacity following orthotopic transplantation. Two out of 68 oocytes obtained by follicular aspiration were confirmed OSCs origin, whereas 17 out of 83 from microdissection. A mature oocyte originating from OSCs developed into 64-cells stage embryo. CONCLUSIONS: Stem cells from NHP adult ovaries can be transplanted and give rise to new oocytes that fertilize and develop into an embryo, suggesting that these stem cells could be a novel approach to treating infertility.