title: Redifferentiation of dedifferentiated chondrocytes and chondrogenesis of human bone marrow stromal cells via chondrosphere formation with expression profiling by large-scale cDNA analysis.
abstract: Characterization of dedifferentiated chondrocytes (DECs) and mesenchymal stem cells capable of differentiating into chondrocytes is of biological and clinical interest. We isolated DECs and bone marrow stromal cells (BMSCs), H4-1 and H3-4, and demonstrated that the cells started to produce extracellular matrices, such as type II collagen and aggrecan, at an early stage of chondrosphere formation. Furthermore, cDNA sequencing of cDNA libraries constricted by the oligocapping method was performed to analyze difference in mRNA expression profiling between DECs and marrow stromal cells. Upon redifferentiation of DECs, cartilage-related extracellular matrix genes, such as those encoding leucine-rich small proteoglycans, cartilage oligomeric matrix protein, and chitinase 3-like 1 (cartilage glycoprotein-39), were highly expressed. Growth factors such as FGF7 and CTGF were detected at a high frequency in the growth stage of monolayer stromal cultures. By combining the expression profile and flow cytometry, we demonstrated that isolated stromal cells, defined by CD34(-), c-kit(-), and CD140alpha(- or low), have chondrogenic potential. The newly established human mesenchymal cells with expression profiling provide a powerful model for a study of chondrogenic differentiation and further understanding of cartilage regeneration in the means of redifferentiated DECs and BMSCs.
authors.affiliation: National Research Institute for Child Health and Development, Tokyo 157-8535, Japan.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Imabayashi
authors.forename: Taisuke
authors.initials: T
authors.lastname: Mori
authors.forename: Satoshi
authors.initials: S
authors.lastname: Gojo
authors.forename: Tohru
authors.initials: T
authors.lastname: Kiyono
authors.forename: Tomoyasu
authors.initials: T
authors.lastname: Sugiyama
authors.forename: Ryotaro
authors.initials: R
authors.lastname: Irie
authors.forename: Takao
authors.initials: T
authors.lastname: Isogai
authors.forename: Jun-ichi
authors.initials: J
authors.lastname: Hata
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Toyama
authors.forename: Akihiro
authors.initials: A
authors.lastname: Umezawa
mesh_terms: D001854:Bone Marrow Cells; D002454:Cell Differentiation / Q000235:genetics; D019902:Chondrocytes / Q000166:cytology*; D020219:Chondrogenesis / Q000235:genetics*; D018076:DNA, Complementary / Q000032:analysis*; D016326:Extracellular Matrix Proteins / Q000235:genetics; D020869:Gene Expression Profiling; D006133:Growth Substances / Q000235:genetics; D006801:Humans; D012333:RNA, Messenger / Q000032:analysis; D012038:Regeneration / Q000235:genetics; D017154:Stromal Cells / Q000166:cytology*
chemical_list: D018076:DNA, Complementary; D016326:Extracellular Matrix Proteins; D006133:Growth Substances; D012333:RNA, Messenger