Levels of Tektin 2 and CatSper 2 in normozoospermic and oligoasthenozoospermic men and its association with motility, fertilization rate, embryo quality and pregnancy rate.

PURPOSE: To compare the expression profiles of Tektin 2 and CatSper 2 motility proteins in the spermatozoa of normozoospermic and oligoasthenozoospermic men and determine its correlation with sperm motility, fertilization rate, embryo quality and pregnancy rate. METHODS: Tektin 2 and CatSper 2 protein expression was studied using Western Blotting and immunofluorescence. Tektin 2 and CatSper 2 protein levels were quantified by ELISA. RESULTS: Oligoasthenozoospermic men were found to have lower fertilization rates, poor embryo quality and lower pregnancy rates as compared to normozoospermic men. The levels of Tektin 2 and CatSper 2 are significantly lower in spermatozoa of oligoasthenozoospermic men as compared to normozoospermic controls; the levels were also lower in immotile fraction as compared to motile fraction of spermatozoa obtained from normozoospermic individuals. The levels of Tektin 2 and CatSper 2 were higher in individuals demonstrating sperm motility >60 % as compared to sperm motility <30 %. Tektin 2 but not CatSper 2 levels were positively associated with fertilization rate, embryo quality and pregnancy rate. CONCLUSION: Levels of Tektin 2 and CatSper 2 proteins are positively associated with sperm motility parameters. Measurements of Tektin 2 levels can be correlated with the clinical outcome of ICSI.

Derivation and characterization of two genetically unique human embryonic stem cell lines on in-house-derived human feeders.

This study describes the successful derivation of two human embryonic stem (hES) cell lines using 53 frozen and 18 fresh "slow-growing" surplus embryos, obtained from collaborating in vitro fertilization clinics, on in-house-derived human feeder layers. The cell lines have been derived by whole embryo culture followed by further expansion of manually dissected inner cell mass from the surrounding trophoectodermal cells. Immunocytochemical localization of cell surface markers like SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81, staining for alkaline phosphatase and reverse transcriptase polymerase chain reaction (RT-PCR) analysis of pluripotency state markers viz. Oct-4, TERT, Nanog, Rex1, and Sox2 indicate that both cell lines possess typical features of embryonic stem cells. Both cell lines exhibit normal female karyotype after 40 passages in culture. Pluripotent nature of the cell lines was confirmed both in vitro and in vivo. Embryoid bodies, formed in suspension culture, express markers for all three lineages as indicated by RT-PCR analysis for SOX 1 (ectoderm), HAND 1 (mesoderm), AFP (endoderm), and CDX2 (trophoectoderm). Teratoma formed in vivo in severe combined immunodeficient mice revealed cells of all the three embryonic germ layers. Comparison of the STR and human leukocyte antigen profiles of these cell lines with the existing human ES cell lines indicate that they are genetically distinct. The addition of our hES cell lines contributes usefully to the globally restricted repertoire of human ES cell lines.