Human chondrogenic paraxial mesoderm, directed specification and prospective isolation from pluripotent stem cells.
Directed specification and prospective isolation of chondrogenic paraxial mesoderm progeny from human pluripotent stem (PS) cells have not yet been achieved. Here we report the successful generation of KDR(-)PDGFRα(+) progeny expressing paraxial mesoderm genes and the mesendoderm reporter MIXL1-GFP in a chemically defined medium containing the canonical WNT signaling activator, BMP-inhibitor, and the Nodal/Activin/TGFβ signaling controller. Isolated (GFP(+))KDR(-)PDGFRα(+) mesoderm cells were sensitive to sequential addition of the three chondrogenic factors PDGF, TGFβ and BMP. Under these conditions, the cells showed robust chondrogenic activity in micromass culture, and generated a hyaline-like translucent cartilage particle in serum-free medium. In contrast, both STRO1(+) mesenchymal stem/stromal cells from adult human marrow and mesenchymal cells spontaneously arising from hPS cells showed a relatively weaker chondrogenic response in vitro, and formed more of the fibrotic cartilage particles. Thus, hPS cell-derived KDR(-)PDGFRα(+ )paraxial mesoderm-like cells have potential in engineered cartilage formation and cartilage repair.
Brown Foundation Institute of Molecular Medicine, University of Texas Health Science Center, Houston, TX 77030, USA.
Katsutsugu
K
Umeda
Jiangang
J
Zhao
Paul
P
Simmons
Edouard
E
Stanley
Andrew
A
Elefanty
Naoki
N
Nakayama
D000328:Adult; D000818:Animals; D000954:Antigens, Surface / Q000235:genetics / Q000378:metabolism; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism; D020219:Chondrogenesis; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008648:Mesoderm / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D011446:Prospective Studies; D020796:Receptor, Platelet-Derived Growth Factor alpha / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism
D000954:Antigens, Surface; D051792:Basic Helix-Loop-Helix Proteins; D019485:Bone Morphogenetic Proteins; D049452:Green Fluorescent Proteins; D018398:Homeodomain Proteins; D010982:Platelet-Derived Growth Factor; D020796:Receptor, Platelet-Derived Growth Factor alpha; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; D040301:Vascular Endothelial Growth Factor Receptor-2; C525671:MEOX1 protein, human; C499230:MIXL1 protein, human; C525591:STRO-1 antigen, human; C513109:TCF15 protein, human