Porcine embryonic stem cells forming germ line chimeras

1)用常规和新型体外成熟/受精系统生产猪1-细胞胚胎，并在添加不同浓度透明质酸的NCSU-23中培养。结果表明，不同的卵母细胞成熟/受精条件下，透明质酸对猪体外胚胎发育的影响不同。2）采用iVMFC法从猪未成熟卵母细胞中获得囊胚，并在STO饲养层上进行培养，以确定其是否可用于ES细胞建立的研究。当转移通过在补充有FCS的培养基中培养完整胚泡而获得的孵化胚泡时，它们附着于饲养细胞。本研究结果表明，猪体外囊胚可用于ES细胞的建立研究。3）我们的研究结果表明，在ES细胞集落中存在单个ES细胞，这些ES细胞保持着高度的全能性或多能性，如果条件允许，单个ES细胞可以与8细胞期胚胎的卵裂球结合，几乎完全是生殖细胞，并伴随泌尿生殖细胞谱系。

1) Porcine 1-cell embryos were produced by the conventional or the new in vitro maturation/fertilization system and cultured in NCSU-23 supplemented with hyaluronic acid at various concentrations. Our results indicate that effect of hysluronic acid on development of in vitro produced porcine embryos varies with the conditions of maturation/fertilization of the oocytes.2) Porcine blastocysts were produced from immature oocytes by using _iVMFC,and cultured on STO feeder layrs to determine whether they can be used for studies on establishment of ES cells. when hatched blastocysts that had been obtained by culturing the intact blastocysts in the medium supplemented with FCS were transferred, they attached to the feeder cells. The results of the present study suggest that in vitro produced porcine blastocysts can be used for studies on establishment of ES cells.3) Our results demonstrated that single ES cells which maintain totipotency or pluripotency of high degree are present in a colony of ES cells, and that single ES cells cojoined with the blastomeres of 8-cell-stage embryos may colonize, if the circumustances allow, almost exclusively the germ cells and concomitantly the urogenital cell lineages.