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Fibrochondrogenesis of hESCs: growth factor combinations and cocultures. Stem Cells Dev 2009 Mar;18(2):283-92

Date

05/06/2008

Pubmed ID

18454697

Pubmed Central ID

PMC3132948

DOI

10.1089/scd.2008.0024

Scopus ID

2-s2.0-62549085191   54 Citations

Abstract

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.

Author List

Hoben GM, Willard VP, Athanasiou KA

Author

Gwendolyn M B Hoben MD Assistant Professor in the Plastic Surgery department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Animals
Biomarkers
Bone Morphogenetic Protein 4
Cell Differentiation
Cell Membrane
Chondrocytes
Chondrogenesis
Coculture Techniques
Collagen
Embryonic Stem Cells
Fibroblasts
Flow Cytometry
Glycosaminoglycans
Humans
Immunohistochemistry
Intercellular Signaling Peptides and Proteins
Mice
Platelet-Derived Growth Factor
Transforming Growth Factor beta3
jenkins-FCD Prod-482 91ad8a360b6da540234915ea01ff80e38bfdb40a