In vitro derivation of chondrogenic cells from human embryonic stem cells.
Human embryonic stem cells (hESCs) have the ability to self-renew and differentiate into any cell lineage of the three germ layers, therefore holding great promise for regenerative medicine applications. However, directing lineage-restricted differentiation of hESCs and obtaining a homogenous differentiated cell population is still a challenge. We previously described a micromass culture system as a model system to study chondrogenic commitment of the hESCs. Using this system, various growth factors including BMP2 and TGFbeta1 direct chondrogenic differentiation and modulate cartilage-specific matrix gene expression in a distinctive manner. Furthermore, a high percentage of differentiated cells exhibit typical morphological characteristics of chondrocytes and express cartilage matrix proteins such as type II collagen and proteoglycans. Chondrogenic cells can be further isolated and cultured to form functional cartilage tissue in vitro. Here, we describe in detail our established protocols to analyze chondrogenic differentiation of hESCs, and possible isolation of chondrogenic cells to form functional cartilaginous tissue.
Toh
Wei Seong
WS
Stem Cell Laboratory, Faculty of Dentistry, National University of Singapore, Singapore.
Lee
Eng Hin
EH
Richards
Mark
M
Cao
Tong
T
D001483:Base Sequence; D055396:Bone Morphogenetic Protein 2 / Q000008:administration & dosage; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002469:Cell Separation; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D020219:Chondrogenesis / Q000187:drug effects; D024042:Collagen Type I / Q000378:metabolism; D024043:Collagen Type II / Q000378:metabolism; D003470:Culture Media; D017931:DNA Primers / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D015870:Gene Expression; D005819:Genetic Markers; D006801:Humans; D007150:Immunohistochemistry; D008954:Models, Biological; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000008:administration & dosage; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053773:Transforming Growth Factor beta1 / Q000008:administration & dosage
C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D024042:Collagen Type I; D024043:Collagen Type II; D003470:Culture Media; D017931:DNA Primers; D005819:Genetic Markers; D012333:RNA, Messenger; D011994:Recombinant Proteins; D053773:Transforming Growth Factor beta1