Cell-cell and cell-extracellular matrix interactions regulate embryonic stem cell differentiation

Cell interactions with the extracellular matrix (ECM) play a critical role in their physiology. Here, we sought to determine the role of exogenous and endogenous ECM in the differentiation of nonhuman primate ESCs. We evaluated cell differentiation from expression of lineage gene mRNA and proteins u...

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Veröffentlicht in:Stem cells (Dayton, Ohio) Ohio), 2007-03, Vol.25 (3), p.553-561
Hauptverfasser: Chen, Silvia S, Fitzgerald, Wendy, Zimmerberg, Joshua, Kleinman, Hynda K, Margolis, Leonid
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container_issue 3
container_start_page 553
container_title Stem cells (Dayton, Ohio)
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creator Chen, Silvia S
Fitzgerald, Wendy
Zimmerberg, Joshua
Kleinman, Hynda K
Margolis, Leonid
description Cell interactions with the extracellular matrix (ECM) play a critical role in their physiology. Here, we sought to determine the role of exogenous and endogenous ECM in the differentiation of nonhuman primate ESCs. We evaluated cell differentiation from expression of lineage gene mRNA and proteins using real-time polymerase chain reaction and immunohistochemistry. We found that ESCs that attached to and spread upon highly adhesive collagen do not differentiate efficiently, whereas on the less adhesive Matrigel, ESCs form aggregates and differentiate along mesoderm and especially endoderm lineages. To further decrease ESC attachment to the substrate, we cultured them either on nonadhesive agarose or in suspension. In both cases, ESCs formed aggregates and efficiently differentiated along endoderm and mesoderm lineages, most strikingly into cardiomyocytes. Aggregates formed by thus-differentiated ESCs started to beat with a frequency of 50-100 beats per minute and continued to beat for approximately a month. In spite of the presence of exogenous ECM, ESCs were dependent on endogenous ECM for their survival and differentiation, as the inhibition of endogenous collagen induced a gradual loss of ESCs and neither a simple matrix, such as type I collagen, nor the complex matrix Matrigel was able to rescue these cells. In conclusion, adhesiveness to various ECM and nonbiological substrates determines the differentiation of ESCs in such a way that efficient cell-cell aggregation, together with less efficient cell attachment and spreading, results in more efficient cell differentiation.
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source Oxford University Press Journals All Titles (1996-Current); MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection
subjects Animals
Cell Communication
Cell Culture Techniques
Cell Differentiation
Cell Division
Collagen
Drug Combinations
Embryonic Stem Cells - cytology
Embryonic Stem Cells - physiology
Extracellular Matrix - physiology
Gene Expression Regulation
Immunohistochemistry
Laminin
Mice
Primates
Proteoglycans
Reverse Transcriptase Polymerase Chain Reaction
title Cell-cell and cell-extracellular matrix interactions regulate embryonic stem cell differentiation
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