Copp A J, Roberts H M, Polani P E
J Embryol Exp Morphol. 1986 Jun;95:95-115.
A microsurgical grafting technique has been used to introduce primordial germ cell (PGC) precursors into intact primitive-streak-stage mouse embryos in vitro. Operated embryos were cultured for 36-40 h and then analysed by a combined histochemical and autoradiographic method. PGC chimaerism occurred in embryos that received grafts of caudal primitive streak cells but not adjacent embryonic endoderm or anterolateral ectoderm/mesoderm cells. Graft-derived PGCs were found to be migrating through the gut endoderm alongside host-derived PGCs in approximately half of the chimaeric embryos whereas in the other 50% of cases PGCs remained at the site of grafting in association with graft-derived somatic cells. A similar pattern of somatic chimaerism was produced by primitive streak and anterolateral ectoderm/mesoderm grafts: the allantois was colonized predominantly, with, in addition, formation of amnion, surface ectoderm and caudal mesoderm in a few embryos. The majority of embryonic endoderm grafts failed to incorporate into host embryos and formed yolk-sac-like vesicles. The findings of this study indicate that PGCs originate from the embryonic ectoderm via the primitive streak during development of the mouse embryo, and anterolateral ectoderm and mesoderm cells are unable to form PGCs after heterotopic grafting to the posterior primitive streak site. The combined microsurgical and embryo culture methods provide an experimental system for the analysis of PGC development in intact mouse embryos.
一种显微外科移植技术已被用于在体外将原始生殖细胞(PGC)前体引入完整的原条期小鼠胚胎。对手术操作后的胚胎进行36 - 40小时的培养,然后通过组织化学和放射自显影相结合的方法进行分析。在接受尾侧原条细胞移植的胚胎中出现了PGC嵌合体,但接受相邻胚胎内胚层或前外侧外胚层/中胚层细胞移植的胚胎未出现。在大约一半的嵌合胚胎中,发现移植来源的PGC与宿主来源的PGC一起穿过肠内胚层迁移,而在另外50%的情况下,PGC与移植来源的体细胞一起留在移植部位。原条和前外侧外胚层/中胚层移植产生了类似的体细胞嵌合模式:尿囊主要被定植,此外,少数胚胎中还形成了羊膜、表面外胚层和尾侧中胚层。大多数胚胎内胚层移植未能整合到宿主胚胎中,而是形成了卵黄囊样囊泡。本研究结果表明,在小鼠胚胎发育过程中,PGC通过原条起源于胚胎外胚层,并且异位移植到后原条部位后,前外侧外胚层和中胚层细胞无法形成PGC。显微外科和胚胎培养相结合的方法为分析完整小鼠胚胎中PGC的发育提供了一个实验系统。