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Lin J.,The Key Laboratory of Animal Biotechnology of Xinjiang | Lin J.,Xinjiang Academy of Animal Science | Wang L.,The Key Laboratory of Animal Biotechnology of Xinjiang | Wang L.,Xinjiang Academy of Animal Science | And 8 more authors.
Asian Journal of Animal and Veterinary Advances | Year: 2012

Oocyte quality directly affects production efficiency of modern assisted reproductive technologies such as the in vitro maturation, fertilization, cultivation of cattle oocytes and sperm injection, embryo transfer and animal cloning. Thus, maternal genes (GDF9, Zarl, Mater and DNMT1) are selected to analyze the relationship between mRNA expressions of these genes and quality of ovine oocyte. This study collects ovine oocytes at GV stage and 12-30 h of in vitro maturation (IVM) respectively, as well as ovine oocytes with the first polar body emitted and not emitted. With the application of quantitative RT-PCR technique, mRNA expression levels of GDF9, Zarl, Mater and DNMT1 of oocytes in the above test groups are tested. It is found that: mRNA relative expression levels of GDF9, Zarl and Mater at GV stage are the highest (p<0.05), while expression of DNMT1 reaches the highest level at 12 h (p<0.05). With the maturity of oocytes, all gene expression levels gradually decline significantly between 18 and 24 h (p<0.05), while the expression levels of four genes present an upward trend at 30 h (p<0.05). Expression of 4 genes in non-polar body oocytes are all higher than those in polar body oocytes (p<0.05). In the natural bare oocytes, expression of each gene is lower than cumulus-oocytes complex. Thus, the test indicates that GDF9, Zarl, Mater and DNMT1 can be the molecular indicators to determine the quality of ovine oocytes. © 2012 Academic Journals Inc. Source

Zhao Y.,The Key Laboratory of Animal Biotechnology of Xinjiang | Zhao Y.,Xinjiang Academy of Animal Science | Lin J.,The Key Laboratory of Animal Biotechnology of Xinjiang | Lin J.,Xinjiang Academy of Animal Science | And 20 more authors.
Journal of Experimental Zoology Part A: Ecological Genetics and Physiology | Year: 2011

Domestic animal embryonic stem (ES) cells would provide an invaluable research tool for genetic breeding and the production of transgenic animals. Unfortunately, authentic domestic animals ES cells have not been established despite progress made over more than two decades. Here, we show that ovine ES-like cells can be efficiently derived and propagated in a semi-defined medium that contains N2, B27, GSK3 inhibitor (CHIR99021), and basic fibroblast growth factor (bFGF). These ovine ES-like cells had a characteristic three-dimensional appearance, showed a bFGF dose-dependence, expressed specific markers such as alkaline phosphatase (AP), Oct-4, Sox2, Nanog and can be maintained for 30 passages. Moreover, these cells differentiated in vitro into neuronal cells, and formed teratomas containing a variety of different tissues including cartilage and neural tissue when injected into kidney capsules of severe combined immunodeficiency (SCID) mice. But the cell lines fail to contribute to embryonic development upon blastocyst transplantation. To our knowledge, this is the first experiment to use semi-defined medium without feeder-cells to derive ES-like cells from ovine blastocysts, and opens the door to deriving authentic ES cells from domesticated ungulates. © 2011 Wiley Periodicals, Inc., A Wiley Company. Source

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