Document Detail

Fibrochondrogenesis of hESCs: growth factor combinations and cocultures.
MedLine Citation:
PMID:  18454697     Owner:  NLM     Status:  MEDLINE    
The successful differentiation of human embryonic stem cells (hESCs) to fibrochondrocyte-like cells and characterization of these differentiated cells is a critical step toward tissue engineering of musculoskeletal fibrocartilages (e.g., knee meniscus, temporomandibular joint disc, and intervertebral disc). In this study, growth factors and primary cell cocultures were applied to hESC embryoid bodies (EBs) for 3 weeks and evaluated for their effect on the synthesis of critical fibrocartilage matrix components: glycosaminoglycans (GAG) and collagens (types I, II, and VI). Changes in surface markers (CD105, CD44, SSEA, PDGFR alpha) after the differentiation treatments were also analyzed. The study was conducted in three phases: (1) examination of growth factors (TGF-beta 3, BMP-2, BMP-4, BMP-6, PDGF-BB, sonic hedgehog protein); (2) comparison of two cocultures (primary chondrocytes or fibrochondrocytes); and (3) the combination of the most effective growth factor and coculture regimen. TGF-beta 3 with BMP-4 yielded EBs positive for collagens I, II, and VI, with up to 6.7- and 4.8-fold increases in GAG and collagen, respectively. Analysis of cell surface markers showed a significant increase in CD44 with the TGF-beta 3 + BMP-4 treatment compared to the controls. Coculture with fibrochondrocytes resulted in up to a 9.8-fold increase in collagen II production. The combination of the growth factors BMP-4 + TGF-beta 3 with the fibrochondrocyte coculture led to an increase in cell proliferation and GAG production compared to either treatment alone. This study determined two powerful treatments for inducing fibrocartilaginous differentiation of hESCs and provides a foundation for using flow cytometry to purify these differentiated cells.
Gwendolyn M Hoben; Vincent P Willard; Kyriacos A Athanasiou
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  Stem cells and development     Volume:  18     ISSN:  1557-8534     ISO Abbreviation:  Stem Cells Dev.     Publication Date:  2009 Mar 
Date Detail:
Created Date:  2009-03-16     Completed Date:  2009-04-08     Revised Date:  2011-08-01    
Medline Journal Info:
Nlm Unique ID:  101197107     Medline TA:  Stem Cells Dev     Country:  United States    
Other Details:
Languages:  eng     Pagination:  283-92     Citation Subset:  IM    
Department of Bioengineering, Rice University, Houston, Texas 77005, USA.
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MeSH Terms
Biological Markers / metabolism
Bone Morphogenetic Protein 4 / pharmacology
Cell Differentiation / drug effects
Cell Membrane / drug effects,  metabolism
Chondrocytes / cytology,  drug effects
Chondrogenesis / drug effects*
Coculture Techniques
Collagen / metabolism
Embryonic Stem Cells / cytology*,  drug effects*
Fibroblasts / cytology*,  drug effects*
Flow Cytometry
Glycosaminoglycans / metabolism
Intercellular Signaling Peptides and Proteins / pharmacology*
Platelet-Derived Growth Factor / pharmacology
Transforming Growth Factor beta3 / pharmacology
Grant Support
Reg. No./Substance:
0/Biological Markers; 0/Bone Morphogenetic Protein 4; 0/Glycosaminoglycans; 0/Intercellular Signaling Peptides and Proteins; 0/Platelet-Derived Growth Factor; 0/Transforming Growth Factor beta3; 9007-34-5/Collagen

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