title: Establishment of bone morphogenetic protein 2 responsive chondrogenic cell line.
abstract: A clonal cell line named RMD-1 was established from the skeletal muscle of a 20-day fetal rat. RMD-1 represents a morphologically homogeneous population of undifferentiated mesenchymal cells, expressing alpha-smooth muscle actin and type I collagen, but no cartilage-associated genes. When cultured in agarose gel containing 100 ng/ml of recombinant human bone morphogenetic protein 2 (rhBMP-2; BMP-2), RMD-1 cells formed colonies and showed chondrocyte-like features as assessed by their ultrastructure, metachromatic staining with toluidine blue, and the production of large hydrodynamic-size proteoglycans. RMD-1 cells also differentiated into chondrocytes when the cells were plated at high density (over 2.5 x 10(5) cells/cm2) on type I collagen and incubated in medium containing 0.5% fetal bovine serum and 100 ng/ml of BMP-2. This chondrogenic differentiation was evidenced by a distinct morphological change into spherical cells, an increase in the levels of sulfated glycosaminoglycans, a decrease in type I collagen mRNA and the expression of cartilage-associated genes, including type II collagen, type IX collagen, aggrecan and alkaline phosphatase. In the presence of ascorbic acid and 10% serum, RMD-1 cells increased in size and expressed type X collagen as well as high alkaline phosphatase activity, then induced matrix mineralization. Thus, RMD-1 is a unique cell line that can differentiate from undifferentiated mesenchymal cells into hypertrophic chondrocytes.
authors.affiliation: First Department of Oral and Maxillofacial Surgery, Osaka University Faculty of Dentistry, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Aikawa
authors.forename: K
authors.initials: K
authors.lastname: Shirasuna
authors.forename: M
authors.initials: M
authors.lastname: Iwamoto
authors.forename: K
authors.initials: K
authors.lastname: Watatani
authors.forename: T
authors.initials: T
authors.lastname: Nakamura
authors.forename: M
authors.initials: M
authors.lastname: Okura
authors.forename: H
authors.initials: H
authors.lastname: Yoshioka
authors.forename: T
authors.initials: T
authors.lastname: Matsuya
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D053674:Aggrecans; D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000378:metabolism*; D002356:Cartilage / Q000166:cytology* / Q000196:embryology; D002417:Cattle; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D002478:Cells, Cultured; D011508:Chondroitin Sulfate Proteoglycans / Q000235:genetics / Q000378:metabolism; D003094:Collagen / Q000235:genetics / Q000378:metabolism; D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans / Q000096:biosynthesis; D006133:Growth Substances / Q000378:metabolism*; D006801:Humans; D037181:Lectins, C-Type; D008648:Mesoderm / Q000166:cytology; D018482:Muscle, Skeletal / Q000166:cytology / Q000196:embryology; D009130:Muscle, Smooth / Q000166:cytology / Q000378:metabolism; D009363:Neoplasm Proteins / Q000378:metabolism*; D011509:Proteoglycans / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism*
chemical_list: C508321:Acan protein, rat; D000199:Actins; D053674:Aggrecans; C525718:BMP2 protein, human; C525720:Bmp2 protein, rat; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D011508:Chondroitin Sulfate Proteoglycans; D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans; D006133:Growth Substances; D037181:Lectins, C-Type; D009363:Neoplasm Proteins; D011509:Proteoglycans; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C499469:recombinant human bone morphogenetic protein-2; D003094:Collagen; D000469:Alkaline Phosphatase