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Establishment of bone morphogenetic protein 2 responsive chondrogenic cell line
Author(s) -
Aikawa Tomonao,
Shirasuna Kanemitsu,
Iwamoto Masahiro,
Watatani Kazuya,
Nakamura Takashi,
Okura Masaya,
Yoshioka Hideo,
Matsuya Tokuzo
Publication year - 1996
Publication title -
journal of bone and mineral research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.882
H-Index - 241
eISSN - 1523-4681
pISSN - 0884-0431
DOI - 10.1002/jbmr.5650110416
Subject(s) - alkaline phosphatase , chondrocyte , chondrogenesis , type ii collagen , aggrecan , mesenchymal stem cell , microbiology and biotechnology , collagen, type i, alpha 1 , chemistry , type i collagen , cartilage , biology , extracellular matrix , biochemistry , endocrinology , anatomy , pathology , osteoarthritis , medicine , articular cartilage , alternative medicine , enzyme
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 α‐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 × 10 5 cells/cm 2 ) 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.