title: Blood island formation in attached cultures of murine embryonic stem cells.
abstract: Differentiation of murine embryonic stem cells in suspension culture results in the formation of cystic embryoid bodies that develop blood islands. In this study pre-cystic embryoid bodies were attached to a substratum, and the program of differentiation was monitored. The attached ES cell cultures formed blood islands on a cell layer that migrated out from the center of attachment and beneath a mesothelial-like cell layer. Morphological and in situ marker analysis showed benzidine-positive hematopoietic cells surrounded by vascular endothelial cells that expressed PECAM and took up DiI-Ac-LDL. Waves of morphological differentiation were evident, suggesting a graded response to differentiation signals. Electron microscopy of the blood islands showed that they were similar to blood islands of cystic embryoid bodies and mouse yolk sacs, and cell-cell junctions were evident among the blood island cells. RNA expression analysis was consistent with the presence of hematopoietic precursor cells of several lineages and a primitive vascular endothelium in the cultures. Thus a program of vascular and hematopoietic development can be elaborated in attached ES cell cultures, and these blood islands are accessible to experimental manipulation.
authors.affiliation: Department of Biology, University of North Carolina at Chapel Hill 27599, USA.
authors.forename: V L
authors.initials: VL
authors.lastname: Bautch
authors.forename: W L
authors.initials: WL
authors.lastname: Stanford
authors.forename: R
authors.initials: R
authors.lastname: Rapoport
authors.forename: S
authors.initials: S
authors.lastname: Russell
authors.forename: R S
authors.initials: RS
authors.lastname: Byrum
authors.forename: T A
authors.initials: TA
authors.lastname: Futch
mesh_terms: D000818:Animals; D015214:Antigens, Differentiation, Myelomonocytic / Q000378:metabolism; D001808:Blood Vessels / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D004730:Endothelium, Vascular / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D008077:Lipoproteins, LDL / Q000378:metabolism; D051379:Mice; D008854:Microscopy, Electron; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism
chemical_list: D015214:Antigens, Differentiation, Myelomonocytic; D015815:Cell Adhesion Molecules; D008077:Lipoproteins, LDL; D019408:Platelet Endothelial Cell Adhesion Molecule-1