Establishment and characterization of tartrate-resistant acid phosphatase and alkaline phosphatase double positive cell lines.
Morphologically macrophage-like cells were cloned from hamster bone marrow cells by coculturing bone marrow cells with hamster chondrocytes. One of the clones (CCP-2) was characterized in the present study. CCP-2 cells were positive in an osteoclast marker enzyme, tartrate-resistant acid phosphatase (TRAP), alkaline phosphatase (ALP) and non-specific esterase (NSE). We showed CCP-2 cells degraded cartilage matrix and hydroxyapatite coated on Osteologic disks. A gelatinase secreted from CCP-2 cells was observed and purified from serum-free conditioned medium of the cells. N-terminal amino acid sequencing of the purified enzyme revealed it was matrix metalloproteinase-9. However, CCP-2 cells failed to express calcitonin receptors, a mature osteoclast marker, even after coculture with osteoblast ST2 cells in the presence of 1alpha, 25-dihydroxyvitamin D3 [1alpha, 25-(OH)2D3]. The cells showed high affinity to types X and I but not to type II collagen. In addition, histochemical studies have shown the presence of tartrate-resistant acid phosphatase and alkaline phosphatase double positive cells at the secondary ossification site of the hamster humerus. From these observations, we concluded that CCP-2 cells are similar to osteoclast but not the same. CCP-2 cells are therefore important tools for investigating chondroclastogenesis/osteoclastogenesis and endochondral ossification.
Masuda
R
R
Division of Biology and Oncology, National Institute of Radiological Sciences, Japan.
Sakiyama
H
H
Nonaka
T
T
Kwan
A
A
Nakagawa
K
K
Moriya
H
H
Imajoh-Ohmi
S
S
Honjo
M
M
Yoshida
K
K
D000135:Acid Phosphatase / Q000032:analysis*; D000469:Alkaline Phosphatase / Q000032:analysis*; D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn; D015415:Biomarkers / Q000032:analysis; D001854:Bone Marrow Cells / Q000166:cytology* / Q000502:physiology; D002448:Cell Adhesion; D002460:Cell Line; D019902:Chondrocytes / Q000502:physiology; D002999:Clone Cells; D018920:Coculture Techniques; D017364:Collagenases / Q000235:genetics / Q000378:metabolism; D006224:Cricetinae; D004792:Enzyme Precursors / Q000235:genetics / Q000378:metabolism; D006132:Growth Plate / Q000033:anatomy & histology / Q000166:cytology / Q000502:physiology; D006801:Humans; D007527:Isoenzymes / Q000032:analysis*; D020780:Matrix Metalloproteinase 9; D008647:Mesocricetus; D051379:Mice; D008969:Molecular Sequence Data; D010010:Osteoclasts / Q000737:chemistry / Q000166:cytology* / Q000201:enzymology; D010012:Osteogenesis; D017386:Sequence Homology, Amino Acid; D000071681:Tartrate-Resistant Acid Phosphatase
D015415:Biomarkers; D004792:Enzyme Precursors; D007527:Isoenzymes; D000469:Alkaline Phosphatase; D000135:Acid Phosphatase; C000605511:Acp5 protein, mouse; D000071681:Tartrate-Resistant Acid Phosphatase; D017364:Collagenases; C120487:pro-matrix metalloproteinase 9; D020780:Matrix Metalloproteinase 9