RRC ID 1210
Author Sakiyama H, Nonaka T, Masuda R, Inoue N, Kuboki Y, Iijima M, Hirabayasi Y, Takahagi M, Yoshida K, Kuriiwa K, Yoshida M, Imajoh-Ohmi S.
Title Characterization of mineral deposits formed in cultures of a hamster tartrate-resistant acid phosphatase (TRAP) and alkaline phosphatase (ALP) double-positive cell line (CCP).
Journal Cell Tissue Res.
Abstract We have established tartrate-resistant acid phosphatase (TRAP) and alkaline phosphatase (ALP) double-positive cell lines (CCP-2, CCP-7, CCP-8) from hamster bone marrow. Accumulation of mineral deposits was observed on the dishes when the clones were cultured in McCoy's 5A medium supplemented with 20% fetal calf serum. The materials were dissolved in 0.05 N HCl, and proteins found in the acid extracts were identified by N-terminal amino acid sequencing. The major components were bovine fetuin and prothrombin precursor. In addition, several cell-derived proteins, such as high mobility group 1 protein (HMG1), secretory leukocyte protease inhibitor (SLPI) and EPV20, a 2.0-kDa milk glycoprotein, were identified. HMG1 was detected, by immunostaining, on the cell surface of all the CCP clones. Metabolically labeled cellular sphingomyelin, sialyllactosylceramide, and proteoglycans were also found in the mineral deposits. Reverse transcription/polymerase chain reaction of CCP-2 mRNA revealed that the cells synthesized alkaline phosphatase, bone sialo protein, and osteonectin, but not matrix Gla protein, osteopontin, and type I collagen. CCP-2 cells formed tumors when injected subcutaneously into nude mice. In the tumor tissue, Alizarin-red-positive nodules surrounded by TRAP- and ALP-positive cells were observed, indicating CCP-2 cells can also induce calcification in vivo.
Volume 309(2)
Pages 269-79
Published 2002-8
DOI 10.1007/s00441-002-0587-y
PMID 12172786
MeSH 3T3 Cells Acid Phosphatase / metabolism* Alkaline Phosphatase / metabolism* Amino Acid Sequence Animals Biomarkers Bone Marrow Cells / cytology Bone Marrow Cells / enzymology* Bone Marrow Cells / ultrastructure Calcification, Physiologic Cell Line* Cricetinae HMGB1 Protein / analysis Injections, Subcutaneous Isoenzymes / metabolism* Kinetics Lipids / analysis Mice Mice, Nude Minerals / chemistry Minerals / metabolism* Osteonectin / biosynthesis Proteoglycans / analysis Tartrate-Resistant Acid Phosphatase
IF 3.043
Times Cited 3
Human and Animal Cells CCP-2 CCP-7 CCP-8 ST2 MC3T3-E1 Nil2C1