NDLI: Differentiation of embryonic stem cells into fibroblast-like cells in three-dimensional type I collagen gel cultures

Content Provider	Springer Nature Link
Author	Togo, Shinsaku Sato, Tadashi Sugiura, Hisatoshi Wang, Xingqi Basma, Hesham Nelson, Amy Liu, Xiangde Bargar, Tom W. Sharp, John G. Rennard, Stephen I.
Copyright Year	2010
Abstract	Fibroblasts are heterogeneous mesenchymal cells that play important roles in the production and maintenance of extracellular matrix. Although their heterogeneity is recognized, progenitor progeny relationships among fibroblasts and the factors that control fibroblast differentiation are poorly defined. The current study was designed to develop a reliable method that would permit in vitro differentiation of fibroblast-like cells from human and murine embryonic stem cells (ESCs). Undifferentiated ESCs were differentiated into embryoid bodies (EBs) with differentiation media. EBs were then cast into type I collagen gels and cultured for 21 d with basal media. The spindle-shaped cells that subsequently grew from the EBs were released from the gels and subsequently cultured as monolayers in basal media supplemented with serum. Differentiated cells showed a characteristic spindle-shaped morphology and had ultrastructural features consistent with fibroblasts. Immunocytochemistry showed positive staining for vimentin and alpha-smooth muscle actin but was negative for stage-specific embryonic antigens and cytokeratins. Assays of fibroblast function, including proliferation, chemotaxis, and contraction of collagen gels demonstrated that the differentiated cells, derived from both human and murine ESCs, responded to transforming growth factor-β1 and prostaglandin E$_{2}$ as would be expected of fibroblasts, functions not expected of endothelial or epithelial cells. The current study demonstrates that cells with the morphologic and functional features of fibroblasts can be reliably derived from human and murine ESCs. This methodology provides a means to investigate and define the mechanisms that regulate fibroblast differentiation.
Starting Page	114
Ending Page	124
Page Count	11
File Format	PDF
ISSN	10712690
Journal	In Vitro Cellular & Developmental Biology - Animal
Volume Number	47
Issue Number	2
e-ISSN	1543706X
Language	English
Publisher	Springer-Verlag
Publisher Date	2010-11-25
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Embryonic stem cell differentiation Fibroblasts Three-dimensional collagen gel cultures Cell Biology Stem Cells Animal Genetics and Genomics Cell Culture Developmental Biology
Content Type	Text
Resource Type	Article
Subject	Cell Biology Developmental Biology Medicine

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