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Embryonic stem cells are a class of any kind in the body cells in vitro infinite self-replication and differentiation of pluripotent cells. Currently, the accepted totipotent embryonic stem cell criteria including: after in vitro and in vivo to the three germ layers of cell differentiation and diploid chimeric into the genital ridge. Kaufman MH 1981 separation of mouse embryonic stem cells so far, step one, to meet these standards only in mice, rats and chickens embryonic stem cell lines. Pig is an important human disease model animals, drugs experimental model animals, at the same time or human xenogeneic organ transplantation the main candidate for the body, its embryonic stem cell lines of the establishment also has been subject to a wide range of concerns, but so far true of pig embryonic stem cell lines (ie satisfies the above criteria) has not been established. In this study, first improved method established mouse embryonic stem cell lines, and mouse embryonic stem cell lines to establish technical system as the basis, through the study on pigs early embryonic development, the pig embryo stem cells of establishing a technical as well as the separation to get real porcine embryonic stem cells possibility. The main experimental methods and results are as follows: the first part KOSR embryonic stem cells cultured The liquid with FBS embryonic stem cell culture medium as the way the lines of of ICR mice ESC separation built. The results show that the method of establishing this improvement can not only isolated blastocysts of ICR mice embryonic stem cell lines, and Construction Department of the efficiency up to 71.4%. Pluripotent embryonic stem cell lines obtained in a detection, the experimental data indicate that the cell lines meet the standard of identity of the mouse embryonic stem cells. The second part of our early embryos of pigs were collected, morphology, diameter measurement experiments. Experiments, we collected data on pig early embryonic development, statistics show that positive contact, the pig early embryonic gestational age and diameter; of developmental synchronization between the same batch of embryos relatively common phenomenon. with a gestational age of the pig early embryonic division general is unscientific. The third part, we try to try to pigs, the in vitro embryo separation embryonic stem cells. The experimental design of three groups of culture fluid for comparison: KOSR embryonic stem cell culture medium, the FBS-embryonic stem cell culture medium, N2/B27 embryonic stem cell culture medium. The statistical results show, KOSR culture solution more suitable for in vivo embryo separation embryonic stem cells. Optimized culture system, we have been a colony of human embryonic stem cell-like, and passed down three generations.
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