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Blood island formation in attached cultures of murine embryonic stem cells
Author(s) -
Bautch Victoria L.,
Stanford William L.,
Rapoport Rebecca,
Russell Scott,
Byrum Robert S.,
Futch Tracy A.
Publication year - 1996
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/(sici)1097-0177(199601)205:1<1::aid-aja1>3.0.co;2-m
Subject(s) - embryoid body , biology , hemangioblast , microbiology and biotechnology , embryonic stem cell , haematopoiesis , stem cell , yolk sac , cellular differentiation , endothelial stem cell , blood cell , immunology , embryo , adult stem cell , genetics , gene , in vitro
Differentiation of murine embryonic stem cells in suspension culture results in the formationof cystic embryoid bodies that develop blood islands. In this study pre‐cystic embryoid bodieswere attached to a substratum, and the program of differentiation was monitored. The attachedES cell cultures formed blood islands on a cell layer that migrated out from the center ofattachment and beneath a mesothelial‐like cell layer. Morphological and in situ marker analysisshowed benzidine‐positive hematopoietic cells surrounded by vascular endothelial cells thatexpressed PECAM and took up DiI‐Ac‐LDL. Waves of morphological differentiation wereevident, suggesting a graded response to differentiation signals. Electron microscopy of the bloodislands showed that they were similar to blood islands of cystic embryoid bodies and mouse yolksacs, and cell‐cell junctions were evident among the blood island cells. RNA expression analysiswas consistent with the presence of hematopoietic precursor cells of several lineages and aprimitive vascular endothelium in the cultures. Thus a program of vascular and hematopoieticdevelopment can be elaborated in attached ES cell cultures, and these blood islands are accessibleto experimental manipulation. © 1996 Wiley‐Liss, Inc.