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Multilineage differentiation of rhesus monkey embryonic stem cells in three-dimensional culture systemsIn the course of normal embryogenesis, embryonic stem (ES) cells differentiate along different lineages in the context of complex three-dimensional (3D) tissue structures. In order to study this phenomenon in vitro under controlled conditions, 3D culture systems are necessary. Here, we studied in vitro differentiation of rhesus monkey ES cells in 3D collagen matrixes (collagen gels and porous collagen sponges). Differentiation of ES cells in these 3D systems was different from that in monolayers. ES cells differentiated in collagen matrixes into neural, epithelial, and endothelial lineages. The abilities of ES cells to form various structures in two chemically similar but topologically different matrixes were different. In particular, in collagen gels ES cells formed gland-like circular structures, whereas in collagen sponges ES cells were scattered through the matrix or formed aggregates. Soluble factors produced by feeder cells or added to the culture medium facilitated ES cell differentiation into particular lineages. Coculture with fibroblasts in collagen gel facilitated ES cell differentiation into cells of a neural lineage expressing nestin, neural cell adhesion molecule, and class III beta-tubulin. In collagen sponges, keratinocytes facilitated ES cell differentiation into cells of an endothelial lineage expressing factor VIII. Exogenous granulocyte-macrophage colony-stimulating factor further enhanced endothelial differentiation. Thus, both soluble factors and the type of extracellular matrix seem to be critical in directing differentiation of ES cells and the formation of tissue-like structures. Three-dimensional culture systems are a valuable tool for studying the mechanisms of these phenomena.
Document ID
20040100093
Acquisition Source
Legacy CDMS
Document Type
Reprint (Version printed in journal)
Authors
Chen, Silvia S.
(NASA Headquarters Washington, DC United States)
Revoltella, Roberto P.
Papini, Sandra
Michelini, Monica
Fitzgerald, Wendy
Zimmerberg, Joshua
Margolis, Leonid
Date Acquired
August 21, 2013
Publication Date
January 1, 2003
Publication Information
Publication: Stem cells (Dayton, Ohio)
Volume: 21
Issue: 3
ISSN: 1066-5099
Subject Category
Life Sciences (General)
Distribution Limits
Public
Copyright
Other

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