[Herpes simplex virus and cytomegalovirus in male ejaculate: herpes simplex virus is more frequently encountered in idiopathic infertility and correlates with the reduction in sperm parameters].

The present investigation was undertaken to study the detection rate of herpes simplex virus (HSV) and cytomegalovirus (CMV) in the ejaculates of males with infertility and to evaluate the impact of virus infection on the major parameters of sperm. Ejaculates from 808 patients were studied. As compared with apparently healthy individuals, the coupled males with primary infertility were found to have HSV more frequently in both the whole ejaculate (31% versus 17%; p = 0.049) and the fraction of actively motile spermatozoa (30% versus 8%; p = 0.016). Ejaculate HSV detection directly correlated with the reduced amount of actively motile spermatozoa (p = 0.0001) and the smaller proportion of morphologically normal forms of germ cells (p = 0.002). CMV was found to have no impact on the motility and morphology of spermatozoids in the ejaculate. Both HSV and CMV in the male ejaculate were significantly more frequently detectable in winter months. The findings lead to the conclusion that HSV is one of the factors for male infertility and can negatively affect the results of assisted reproductive technologies.

[Isolation and long-term cultivation of rabbit embryonic stem cells]. / Polychenie i dlitel'noe kul'tivirovanie émbrional'nykh stvolovykh kletok krolika.

A cell line similar to embryonic stem cells has been isolated from 5-day rabbit embryos. After the disruption of zona pellucida, embryos were mechanically separated into cel aggregates, which were then cultivated on a feeder layer of primary mouse embryonic fibroblasts. Cells similar to embryonic stem cells had high nuclear-to-cytoplasm ratio. It has been demonstrated for the first time that the inner cell mass and embryonic stem cells of rabbits expressed the activity of alkaline phosphatase. Between 10 and 30% of cells retained this expression and pluripotent characteristics up to the 23rd transfer. After 14-17 transfers the obtained embryonic cells could be induced to differentiation in vitro, which resulted in the formation of embryoid bodies. After the 23rd transfer, the line of rabbit embryonic cells was lost as a result of its differentiation.

[The effect of the developmental stage and of the cultivation conditions for cow and mouse embryos on the isolation of ES-like cells]. / Vliianie stadii razvitiia i uslovii kul'tivirovaniia émbrionov korov i myshei na poluchenie ES-podobnykh kletok.

In order to study the effect of the stage of development and culturing conditions on the preparation of ES-like cells, we performed experiments on long-term culturing of cow and mouse embryos. We showed that success in isolation of ES-like cells significantly depends on the stage of development. All ES-like cells of cows and mice maintained in culture over three or more transfers were obtained from the embryos at the stage of the released embryo or the late blastocyst, respectively. We found that isolation of ES-like cells of mice was successful only when alpha-MEM medium was used without the addition of BRL-conditioned medium.

[The factors influencing the isolation of pig embryonic stem cells]. / Faktory, vliiaiushchie na poluchenie émbrional'nykh stvolovykh kletok svinei.

To study factors influencing the preparation of porcine ES cells, intact pig embryos at various stages of preimplantation development, as well as clusters of embryonic cells prepared from embryos at the stage of late or expanded blastocyst were cultivated in alpha-MEM, alpha-MEM + 50% BRL-conditioned medium, or in PBS + 20% FCS. NIH 3T3 cells, or pig granulosa cells were used as a feeder layer. It has been demonstrated that incubation conditions seriously affect development, release, and attachment to the feeder layer of both intact pig embryos and clusters of embryonic cells. However, the stage of preimplantation development is the most important factor, defining success in preparation of pig ES cells. It has been shown that embryoblast of pig embryos at stages of early, middle, late, and expanded blastocyst consists of 4-5, 8-10, 12-15, and 16-18 cells, respectively. This leads us to the conclusion that the rate of proliferation of ICM cells from pig embryos is very low. These results indirectly confirm the existence in embryonic development of pigs of metabolically resting ICM cells up to the beginning of gastrulation.