Porcine embryonic stem cells forming germ line chimeras

1)采用常规和新型体外成熟/受精系统制备猪1细胞胚胎，在添加不同浓度透明质酸的NCSU-23培养基中培养。结果表明，羟丙酸对体外培养的猪胚胎发育的影响随卵母细胞成熟/受精条件的不同而不同。2)利用_iVMFC技术制备猪未成熟卵母细胞囊胚，并在STO饲喂层上培养，以确定其是否可用于胚胎干细胞的建立研究。将完整囊胚在添加了FCS的培养基中培养获得的孵化囊胚转移到饲养细胞上。本研究结果表明，体外培养的猪囊胚可用于胚胎干细胞的构建研究。3)我们的研究结果表明，在胚胎干细胞集落中存在着保持高度全能性或多能性的单个胚胎干细胞，并且在条件允许的情况下，单个胚胎干细胞与8细胞期胚胎的卵裂球结合可以几乎完全定植生殖细胞和伴随的泌尿生殖细胞系。

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.