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Hall V.,Copenhagen University | Hinrichs K.,Texas A&M University | Lazzari G.,Laboratory of Reproductive Technologies | Betts D.H.,University of Western Ontario | Hyttel P.,Copenhagen University
Veterinary Journal | Year: 2013

Over many decades assisted reproductive technologies, including artificial insemination, embryo transfer, in vitro production (IVP) of embryos, cloning by somatic cell nuclear transfer (SCNT), and stem cell culture, have been developed with the aim of refining breeding strategies for improved production and health in animal husbandry. More recently, biomedical applications of these technologies, in particular, SCNT and stem cell culture, have been pursued in domestic mammals in order to create models for human disease and therapy. The following review focuses on presenting important aspects of pre-implantation development in cattle, pigs, horses, and dogs. Biological aspects and impact of assisted reproductive technologies including IVP, SCNT, and culture of pluripotent stem cells are also addressed. © 2013 Elsevier Ltd. Source

Sartori C.,Laboratory of Reproductive Technologies | Didomenico A.I.,Roslin Institute | Thomson A.J.,Roslin Institute | Milne E.,Roslin Institute | And 3 more authors.
Cellular Reprogramming | Year: 2012

Pluripotential stem cells from livestock offer an exciting prospect for the biotechnology industry. Applying strategies established for the derivation of murine induced pluripotential stem cells (iPSCs), we have isolated ovine iPSCs that can give rise to cells characteristic of all three germ cell layers both in vitro from embryoid bodies and in teratomas in vivo. Furthermore, although at a low level, these ovine iPS cells can contribute to live-born chimeric lambs. Colonies derived from ovine embryonic fibroblasts transfected with murine cMyc, Klf4, Oct4, and Sox2 displayed smooth domes with sharp edges when grown in human embryonic stem cell (ESC) medium but not in mouse ESC medium. These ovine iPSCs were alkaline phosphatase positive, expressed Nanog, and had a normal karyotype. These cells represent an important step in the understanding of mechanistic nature of pluripotency in ungulates. © 2012, Mary Ann Liebert, Inc. Source

Dieci C.,Laboratory of Reproductive Technologies | Lodde V.,Laboratory of Reproductive Technologies | Lodde V.,University of Milan | Franciosi F.,University of Milan | And 8 more authors.
Biology of Reproduction | Year: 2013

In the pig, the efficiency of in vitro embryo production and somatic cell nuclear transfer (SCNT) procedures remains limited. It has been suggested that prematuration treatments (pre-IVM) based on the prolongation of a patent, bidirectional crosstalk between the oocyte and the cumulus cells through gap junction mediate communication (GJC), with the maintenance of a proper level of cAMP, could improve the developmental capability of oocytes. The aim of this study was to assess: 1) dose-dependent effects of cilostamide on nuclear maturation kinetics, 2) the relationship between treatments on GJC functionality and large-scale chromatin configuration changes, and 3) the impact of treatments on developmental competence acquisition after parthenogenetic activation (PA) and SCNT. Accordingly, cumulus-oocyte complexes were collected from 3- to 6-mm antral follicles and cultured for 24 h in defined culture medium with or without 1 lM cilostamide. GJC functionality was assessed by Lucifer yellow microinjection, while chromatin configuration was evaluated by fluorescence microscopy after nuclear staining. Cilostamide administration sustained functional coupling for up to 24 h of culture and delayed meiotic resumption, as only 25.6% of cilostamide-treated oocytes reached the pro-metaphase I stage compared to the control (69.7%; P < 0.05). Moreover, progressive chromatin condensation was delayed before meiotic resumption based upon G2/M biomarker phosphoprotein epitope acquisition using immunolocalization. Importantly, cilostamide treatment under these conditions improved oocyte developmental competence, as reflected in higher blastocyst quality after both parthenogenetic activation and SCNT. © 2013 by the Society for the Study of Reproduction, Inc. Source

Lazzari G.,Laboratory of Reproductive Technologies | Colleoni S.,Laboratory of Reproductive Technologies | Galli C.,Laboratory of Reproductive Technologies | Galli C.,University of Bologna
Methods in Molecular Biology | Year: 2013

Induction of neural differentiation from embryonic pluripotent stem cells (ES/EpiSc), both of mouse and primates, has been extensively published by several research teams. However, direct derivation of organized neuroectoderm in vitro from blastocyst stage embryos of rodents or primates has not been reported so far. Here we describe a method of direct neural differentiation from the inner cell mass cells of preimplantation bovine embryos, without the intermediate step of ES/EpiSc cells derivation (Lazzari et al., Stem cells 24:2514-2521, 2006). Proliferating neural precursors cells lines, and both central and peripheral nervous system derivatives, can be obtained providing a unique in vitro model of early neurulation events in mammals. © 2013 Springer Science+Business Media, New York. Source

Busby S.-A.,Glasgow Caledonian University | Crossan C.,Glasgow Caledonian University | Godwin J.,Glasgow Caledonian University | Petersen B.,Friedrich Loeffler Institute | And 5 more authors.
Xenotransplantation | Year: 2013

The hepatitis E virus (HEV) is considered a zoonotic pathogen. In xenotransplantation, given the high prevalence of HEV infection in pigs, the risk of zoonotic transmission from a porcine source is considered high. Currently no clear data are available on how to diagnose and eliminate HEV in herds used for medical purposes and the importance of viral infection at the stage of harvest. In this study, several groups of animals currently used for medical purposes were found RNA positive in both serum and faeces for HEV genotype 3. In addition, viraemia was found in animals up to 3.6 yr of age, which is much longer than originally expected. Herd transmission rates appeared to be significantly lower in animals kept under minimal barrier conditions, compared with those observed for commercial animals, and as expected, segregation of animals at an early age prevented spread of infection. This study makes suggestions to ensure appropriate detection and eradication of HEV from a donor herd to be used for xenotransplantation purposes. © 2013 John Wiley & Sons A/S. Source

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