We have separated for enzyme analysis the following layers that surround the conceptus at midgestation: decidua, trophoblast, parietal endoderm (including Reichert’s membrane), visceral endoderm, yolk-sac mesoderm and amnion. Measurement of several catabolic enzyme activities )N-acetyl-β,D-hexosaminidase, β-glucuronidase, alkaline and acid phosphatases and non-specific esterases) in these tissues indicates that they are biochemically distinct, perhaps reflecting the different functions that they perform in providing the embryo proper with a desirable environment for differentiation and development. Our studies also provide an example of how visceral endoderm cells can effectively block passage of maternal macromolecules (in this case a serum esterase) in the fetal circulation. Finally, since there is often difficulty in distinguishing among early embryonic and extra-embryonic cell types produced in teratocarcinoma cultures, we have considered how our observations might be of use in this respect, particularly in discriminating between visceral and parietal endoderm.

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