The role of cell interactions in the differentiation of teratocarcinoma-derived parietal and visceral endoderm.
Cell interactions have been implicated in the differentiation of visceral and parietal endoderm in the developing mouse embryo. Embryoid bodies formed from F9 embryonal carcinoma cells have been useful in characterizing the events which lead to endoderm formation. As part of our effort to specify the interactions which may be involved in this process we have isolated visceral endoderm-like cells (VE) from F9 embryoid bodies and cultured them under various conditions. Using a combination of immunoprecipitation and enzyme-linked immunosorbent assay, we demonstrate that monolayer culture of these cells on a number of different substrates leads to a dramatic decrease in the level of alphafetoprotein (AFP), a VE-specific marker. Northern blot analysis of AFP mRNA indicates very low levels of this message are present after 48 hr in monolayer culture. Coincident with the drop in AFP levels is an increase in the levels of the cytokeratin Endo C and tissue plasminogen activator, both markers for parietal endoderm (PE). Morphological evidence at the ultrastructural level supports a transition from VE to PE. In contrast, the VE phenotype can be maintained in vitro by interaction with aggregates, but not monolayers, of stem cells. In addition, culturing the cells on the curved surface of gelatin-coated dextran beads, but not on a flat gelatin surface facilitates AFP expression and the cells are morphologically intermediate between VE and PE cells. The potential role of junctional complexes and cell shape are discussed.
Casanova
J E
JE
Department of Biology, Wesleyan University, Middletown, Connecticut 06457.
Grabel
L B
LB
D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002450:Cell Communication; D002454:Cell Differentiation; D004475:Ectoderm / Q000473:pathology; D004707:Endoderm / Q000473:pathology*; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D007163:Immunosorbent Techniques; D007633:Keratins / Q000378:metabolism; D051379:Mice; D008854:Microscopy, Electron; D012333:RNA, Messenger / Q000378:metabolism; D013234:Stem Cells / Q000502:physiology; D013724:Teratoma / Q000473:pathology*; D010959:Tissue Plasminogen Activator / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014781:Viscera; D000509:alpha-Fetoproteins / Q000096:biosynthesis / Q000235:genetics
D012333:RNA, Messenger; D000509:alpha-Fetoproteins; D014212:Tretinoin; D003994:Bucladesine; D007633:Keratins; D010959:Tissue Plasminogen Activator