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Parthenotes as a source of embryonic stem cells
The derivation and study of human embryonic stem cell lines, despite their potential therapeutic usefulness, raise considerable ethical, religious, legal and political concerns because it inevitably leads to the destruction of viable embryos. In an attempt to bridge the division between ethical questions and potential scientific and medical benefits, considerable efforts have been devoted to the search for alternative sources of pluripotent cell lines. In this review we discuss the use of artificial parthenogenesis as a way to create entities, called parthenotes, that may represent an alternative ethical source for pluripotent cell lines. We describe the biological differences between parthenotes and embryos, in order to provide a rationale for the discussion on whether their use can be acceptable as a source of stem cells. We present data derived from animal models on the extent parthenogenetic stem cells are similar to biparental cell lines and discuss these aspects in the context of their extension to the human species. Finally, we present experiments recently carried out in our laboratory that allowed us to generate human parthenotes through artificial activation of human oocytes and to use them as a source for the derivation of parthenogenetic pluripotent cell lines
In vitro maturation of farm animal oocytes : a useful tool for investigating the mechanisms leading to full term development
Due to logistical and economic reasons, assisted reproduction of domestic animals has been based mostly on the use of oocytes isolated from ovaries collected at the slaughterhouse. In order to propagate valuable or rare genetic material, perform somatic cell nuclear transfer or generate genetically modified animals, it is essential to obtain fully competent oocytes that will allow full-term development of the in vitro-produced embryos. Such a need makes clear the crucial role played by oocyte quality. In fact, it is easy to compromise the oocyte's developmental potential but it is impossible to restore once it has been lost. Almost three decades after the first cow, sheep, goat, horse and pig in vitro-generated offspring were born, a large body of information has accumulated on the mechanisms regulating oocyte competence and on how the latter may be preserved during all the required manipulations. The amount of knowledge is far from complete and many laboratories are actively working to further expand it. In this review we will highlight the aspects of the ongoing research in which we have been actively involved
Oocytes and ovarian follicles as targets of endocrine disrupters: consequences for reproductive health
Direct inhibitory effect of somatostatin on the growth of the human prostatic cancer cell line LNCaP: possible mechanism of action
The effects of somatostatin-14 (S) on the proliferation of the human prostatic cancer cell line (LNCaP, lymph node carcinoma of the prostate) and on the amount of proteins secreted by these cells have been studied. LNCaP cells were treated for 3 days with different doses of S. The hormone significantly inhibits cell proliferation at doses comprised between 0.4 and 2 nmol, as indicated by a significant decrease in the incorporation of 3H-thymidine as well as in the uptake of [a-32P]dATP. The direct antiproliferative effect of S is reversible, since cell proliferation returns to normal 4 days after withdrawal of the hormone from the medium. This action of S is not due to cytotoxic side-effects, since no changes in the incorporation of 35S-methionine are observed. Total RNA and protein synthesis do not appear to be modified by the hormone in vitro. The hormone probably exerts its antiproliferative effect on LNCaP cells by stimulating phosphotyrosyl protein phosphatases, since the inhibition of growth induced by S (1 nmol) is reversed by sodium vanadate (2 and 4 μmol), a potent inhibitor of the process of tyrosine dephosphorilation. In addition, S (1 nmol/L) induces a significant decrease in the amounts of proteins secreted by LNCaP cells. Also the antisecretory action of S seems to be mediated by the activation of phosphotyrosyl protein phosphatases, since sodium vanadate is able to reverse the action of S on this parameter. In conclusion, the present data suggest for the first time that S exerts a direct inhibitory effect on LNCaP cell proliferation and protein secretion, two effects probably mediated by the activation of phosphotyrosyl protein phosphatases
No shortcuts to pig embryonic stem cells
The establishment of embryonic stem cell (ESC) lines in domestic species could have great impact in the agricultural as well as in the biomedical field. In particular, derivation of pig ESC would find important applications aimed at improving health and production traits of this species through genetic engineering. Similarly, the immunological, morphological, physiological, and functional similarities to the human make the pig a very effective and suitable animal model for biomedical studies and pre-clinical trials. While proven blastocyst-derived mouse and human ESC lines have been established, no validated porcine ESC (pESC) lines are available. In the present manuscript we briefly discuss some of the factors that make the establishment of ESC lines in the pig, and in animal species other than mouse and human, a very slow process. The paucity of information related to morphology, pluripotency markers, differentiation capability hampers a thorough evaluation of the validity of putative lines. These difficulties are further increased by the lack of reliable antibodies, reagents, and in vitro culture systems that could ensure reliable results in the pig and allow for the screening and long-term maintenance of pESC. Data from the literature suggest that similar regulatory pathways are likely to exist among different species. Coupling of these pathways with their distinct expression patterns, the relative concentrations of pluripotency-related molecules, and timing of embryo development, along with supportive micro-environmental conditions, would appear to vary in a species-specific manner. We feel that the understanding of these subtle but meaningful diversities may provide beneficial information about the isolation of genuine porcine embryonic stem cells
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