Cell relationships during aggregation between preimplantation embryos and teratocarcinoma-derived cells

Abstract

Cleavage-stage mouse embryos aggregate and form chimaeric blastocysts with embryonal carcinoma (EC) cells. We used scanning and transmission electron microscopy to study cell relationships during aggregate formation between 8-cell-stage embryos and F9 EC cells. Relations between heterotypic cells were similarly studied in aggregation experiments with embryos and teratocarcinoma-derived visceral (PSA5-E) and parietal (PYS-2) endoderm cells and in experiments with EC cells and endoderm cells. The embryos and F9 cells always adhered to each other and rapidly formed compacted aggregates. Numerous microvilli and cell processes, originating from both embryo and EC cells, extended between the two cell types during adhesion and early phases of aggregation. The aggregation process involved spreading of the blastomeres on the EC cells. Frequent adherent junctions and close contacts, including possible focal gap or tight junctions were observed between the embryo and F9 cells after 3 h of culture. Apparent gap or tight junctions were infrequent during the early phases of aggregation but during further culture, extensive typical gap junctions were also seen between embryo and EC cells. The embryos adhered only irregularly and loosely to PSA5-E and PYS-2 cells; this interaction never led to aggregate formation comparable to that seen in the experiments with embryos and EC cells. Close contacts but no gap or tight junctions could be observed between the embryo and endoderm cells. On the other hand, both PSA5-E and PYS-2 cells readily adhered to and aggregated with EC cells. The present results suggest that microvilli and cell processes mediate membrane interactions during adhesion and early phases of aggregation between embryos and EC cells. During aggregation, blastomeres spread over the EC cells, and rapid formation of adherent junctions and close contacts, including possible focal gap or tight junctions is involved during the early phases of this process. After this initial phase, typical gap junctions are also seen between the embryo and EC cells. Interestingly, adhesive properties of embryo and EC cells differ: the former aggregate only with EC cells, whereas the latter do so also with teratocarcinoma-derived visceral and parietal endoderm cells. Mechanisms operating in the morphogenetic movement of cells in this experimental setup may be involved also in the development of the blastocyst in vivo.