Twelve blastocysts, collected 7-12 days after ovulation (Day 0), were examined by light and electron microscopy to investigate the nature of the relationship of the polar trophoblast (Rauber's layer) to the inner cell mass. On Day 7, the polar trophoblast was intact and formed a flattened layer overlying the epiblast cells of the inner cell mass. As blastocysts enlarged to greater than 1 mm in diameter, small discontinuities appeared in the polar trophoblast, where epiblast cells intruded onto the surface. At this time, trophoblast cells adhered closely to adjacent and underlying epiblast cells, forming an irregular layer of cells capping the epiblast. With continued increase in blastocyst size, polar trophoblast cells became isolated but maintained their characteristic apical endocytic structures. By Days 10-12, the scattered trophoblast cells showed evidence of deterioration, and vacuoles containing cell debris were common within the epiblast. It is suggested that polar trophoblast cells become scattered, rather than withdrawing as a unit, because they become more adherent to subjacent epiblast cells than to adjacent trophoblast cells. It is further suggested that most of the isolated cells are eventually phagocytosed by epiblast cells.

Summary On Day 7 of pregnancy, when the inner cell mass coalesces to one side of the blastocyst, a discontinuous layer of cells is present underlying the inner cell mass and isolated cells were found around the mural trophoblast. As early as Day 8, these cells that were scattered around the blastocyst cavity unite to form a continuous endodermal layer, resulting in a bilaminar blastocyst. The endodermal cells have slightly bulging apical surfaces towards the cavity of the blastocyst, but in many areas have flanges and projections that form complex basolateral compartments within the endoderm and especially between the endoderm and trophoblast. These cells also develop an extensive series of unusual apical ridges. With the development of the primitive streak and the trilaminar blastocyst, angiogenesis proceeds from the forming embryo into the yolk sac. At first the anastomotic vessels form a simple mesh, but by Day 20, erythroblasts are present in the newly formed sinus terminalis. The loss of the capsule (Day 21) and the development of the embryonic circulation establish the relationships of the definitive yolk sac. There is a bilaminar region with endodermal cells showing basolateral compartmentalisation and a specialised sinus terminalis region bordering the choriovitelline placenta. The largest endodermal cells are associated with the choriovitelline placenta, but a substantial cuboidal lining also covers the splanchnopleuric portion of the vascular yolk sac. The allantochorion rapidly assumes predominance in respiratory exchanges, and the choriovitelline placenta not only decreases in relative area but increases in thickness. However, the 3 major regions of the yolk sac persist for several weeks after their formation, and both the vasculature and especially the endodermal cells remain robust despite the decrease in size of the yolk sac. It is concluded that the pleomorphic endodermal cells of the equine yolk sac differentiate in response to local signals. By Day 25 the allantochorion is an increasingly significant exchange unit, and the continued robustness of the vascular portions of the yolk sac and especially its endodermal layer suggests that synthetic and metabolic activities of the endodermal cells will be the dominant function of the yolk sac subsequently.

The interaction of cytotrophoblast with maternal endometrium, especially endometrial blood vessels, was examined in macaque gestational stages between 2 and 8 days after the onset of implantation. Serial sectioning of these early implantation sites allowed immunostaining of consecutive sections with a number of different antibodies, facilitating cell identification. In the earliest implantation site, immunostaining showed that antibody to cytokeratin stained cytotrophoblast, syncytial trophoblast, epithelial plaque and endometrial gland cells. However, only those cytotrophoblast cells near the maternal-fetal border and within vessels showed surface staining for neural cell adhesion molecules and only syncytial trophoblast showed SP1 reactivity. Even at this early stage cytotrophoblast filled the lumen of superficial arterioles, whereas dilated venules contained only a few cytotrophoblast cells. In later stages endovascular cytotrophoblast not only plugged many spiral arterioles but also migrated into the walls of these arterioles, and progressed into deeper coils. Displacement of endothelial cells and disruption of vessel walls were illustrated with antibody to factor VIII, TGFoc, and desmin. Clusters of cytotrophoblast cells at the fetal-maternal interface tended to bypass clusters of epithelial plaque cells and larger clusters of maternal fibroblasts, but readily entered all vascular spaces. Consequently the vascular system constituted a major pathway of invasion, although the arterioles were the only component substantially invaded beyond the trophoblastic-shell/endometrial border.

No abstract

During the blastocyst stage of development in the baboon, the inner cell mass changes from an irregular accumulation of cells within the cavity of the blastocyst to a disk at one side of the blastocyst and finally to a spherical mass of epiblast cells exhibiting a distinct polarity. The cells that will become the primitive endoderm are first seen as flattened but undifferentiated cells on the cavity side of the disk-shaped inner cell mass. After endoderm cells develop their typical cytological characteristics, they extend well beyond the inner cell mass to form parietal endoderm. A basal lamina develops associated with the epiblast cells and mural trophoblast, but not with either parietal or visceral endoderm. Cytological differentiation of inner cell mass cells includes increased numbers of polyribosomes and a change in mitochondria from long, convoluted structures to short, more typical shapes. Evidence that epiblast is polarized is seen by the late zonal blastocyst stage. Apical junctional complexes develop within the center of the epiblast. These junctions presage the development of the potential amniotic cavity. Large vacuoles containing cell debris, some of which contain nuclear fragments, are present at all stages. Extensive cell death occurs during growth of the blastocyst, but the pattern appears to be random and products of cell death are readily phagocytized by adjacent cells.