Effects of embryotrophic factors on the embryogenesis and organogenesis of mouse embryos in vitro.
Effects of embryotrophic factors on the embryogenesis and organogenesis of mouse embryos in vitro.
复制标题
胚胎营养因子对小鼠体外胚胎发生和器官发生的影响。
作者：
I. Ishiwata;Y. Tokieda;K. Kiguchi;K. Sato;H. Ishikawa
为了研究体外胚胎发生和器官发生，用添加10%FCS和胚胎营养因子（ETFs）的α-MEM培养小鼠2细胞胚胎。ETFs从SKG-Ⅱ-SF细胞系的条件培养基中分离，SKG-Ⅱ-SF细胞系来源于人子宫颈表皮样癌。IL-1 β、IL-6、IL-8、EGF、GH、PDGF-AB、碱性FGF、VEGF在该细胞系的条件培养基中也被检测到。使用ETFs和添加10%FCS的培养基，23%的小鼠2细胞期胚胎发育到双层盘期，13%发育到三层芽盘期，9%的胚胎在卵黄囊中观察到血胰岛，7%（28个胚胎）的胚胎观察到心脏搏动。此外，还观察了原始器官，如肝、心、肾、脊索、视网膜样结构等。通常，使用来自两个细胞阶段胚胎的ETF在体外发育与原始条纹阶段（双层胚盘胚胎）相关的结构。这些结构与体内正常胚胎中相同阶段的结构非常相似。在原始条纹期之后，培养的胚胎与正常胚胎的同一时期没有相似之处。这些胚胎的外观都不正常。另一方面，当仅使用10%FCS补充的培养系统时，三层盘阶段胚胎从未发育。
In order to study embryogenesis and organogenesis in vitro, two cell mouse embryos were cultured with alpha-MEM supplemented 10% FCS and embryotrophic factors (ETFs). The ETFs were separated from the conditioned medium of a SKG-II-SF cell line derived from a human uterine cervical epidermoid carcinoma. IL-1 beta, IL-6, IL-8, EGF, GH, PDGF-AB, basic FGF, VEGF were also detected in the conditioned media of this cell line. Using ETFs and a 10% FCS supplemented culture medium, 23% of the mouse two cell stage embryos developed to the bilaminar disc stage, 13% to the trilaminar germ disc stage, 9% were observed with blood islets in the yolk sac, and the heart beat was noted in 7% (28 embryos) of the embryos. Furthermore, primordial organs, such as the liver, heart, kidney, notochord, retina-like structure, etc. were observed. Usually, structures associated with the primordial streak stage (bilaminar germ disc embryo) developed in vitro using ETFs from two cell stage embryos. These closely resemble structures found at the same stage in the normal embryo in vivo. After the primordial streak stage, the cultured embryos showed no resemblance to the same stage in normal embryos. None of the external appearances of these embryos appeared normal. On the other hand, trilaminar disc stage embryos never developed when using only a 10% FCS supplemented culture system.