Department of Veterinary Medicine, FZEA, University of Sao Paulo, Pirassununga, São Paulo, Brazil.
Microsc Res Tech. 2013 Jul;76(7):756-66. doi: 10.1002/jemt.22227. Epub 2013 May 7.
The yolk sac (YS) is the main source of embryonic nutrition during the period when the placenta has not yet formed. It is also responsible for hematopoiesis because the blood cells develop from it as part of the primitive embryonic circulation. The objective of this study was to characterize the transitional area between the YS and primitive gut using the techniques of light microscopy, transmission electron microscopy, and immunohistochemistry to detect populations of pluripotent cells by labeling with Oct4 antibody. In all investigated embryos, serial sections were made to permit the identification of this small, restricted area. We identified the YS connection with the primitive intestine and found that it is composed of many blood islands, which correspond to the vessels covered by vitelline and mesenchymal cells. We identified large numbers of hemangioblasts inside the vessels. The mesenchymal layer was thin and composed of elongated cells, and the vitelline endodermal membrane was composed of large, mono- or binucleated cells. The epithelium of the primitive intestine comprised stratified columnar cells and undifferentiated mesenchymal cells. The transitional area between the YS and the primitive intestine was very thin and composed of cells with irregular shapes, which formed a delicate lumen containing hemangioblasts. In the mesenchyme of the transitional area, there were a considerable number of small vessels containing hemangioblasts. Using Oct4 as a primary antibody, we identified positive cells in the metanephros, primordial gonad, and hepatic parenchyma as well as in YS cells, suggesting that these regions contain populations of pluripotent cells.
卵黄囊(YS)是胎盘尚未形成时胚胎营养的主要来源。它还负责造血,因为血细胞从它发育而来,是原始胚胎循环的一部分。本研究的目的是使用光镜、透射电镜和免疫组织化学技术来描述卵黄囊和原始肠之间的过渡区域,通过 Oct4 抗体标记来检测多能细胞群体。在所有研究的胚胎中,都进行了连续切片以允许识别这个小而受限的区域。我们确定了卵黄囊与原始肠的连接,并发现它由许多血岛组成,这些血岛对应于被卵黄和间充质细胞覆盖的血管。我们在血管内发现了大量的成血管细胞。间充质层很薄,由伸长的细胞组成,卵黄内胚层膜由大的单核或双核细胞组成。原始肠的上皮由分层柱状细胞和未分化的间充质细胞组成。卵黄囊和原始肠之间的过渡区域非常薄,由形状不规则的细胞组成,形成一个包含成血管细胞的精致管腔。在过渡区域的间充质中,有相当数量的含有成血管细胞的小血管。使用 Oct4 作为一抗,我们在中肾、原始性腺和肝实质以及卵黄囊细胞中鉴定出阳性细胞,表明这些区域含有多能细胞群体。