Establishment of an osteoblast-like cell line, MMC2, from p53-deficient mice

• Published in: Bone (New York, N.Y.) Vol. 21; no. 4; pp. 313 - 319
• Main Authors: Nakayama, T., Kanoe, H., Sasaki, M.S., Aizawa, S., Nakamura, T., Toguchida, J.
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier Inc 01-10-1997; Elsevier Science
• Subjects: Alkaline Phosphatase - metabolism; Animal cells; Animals; Ascorbic Acid - pharmacology; Biological and medical sciences; Biotechnology; Bone cell; Bone morphogenetic protein; Bone Morphogenetic Proteins - biosynthesis; Calcification, Physiologic; Cell Differentiation; Cell Division; Cell Line - drug effects; Collagen - metabolism; Differentiation; Establishment of new cell lines, improvement of cultural methods, mass cultures; Eukaryotic cell cultures; Extracellular Matrix - ultrastructure; Fundamental and applied biological sciences. Psychology; Genes, Tumor Suppressor; Immortalized cell line; Methods. Procedures. Technologies; Mice; Mice, Nude; Microscopy, Electron; Osteoblast; Osteoblasts - cytology; Osteoblasts - metabolism; Osteocalcin - metabolism; p53 gene; Phenotype; Skeleton and joints; Skull - cytology; Skull - ultrastructure; Space life sciences; Tumor Suppressor Protein p53 - deficiency; Tumor Suppressor Protein p53 - genetics; Vertebrates: osteoarticular system, musculoskeletal system; Osteoblast; Bone cell; Bone morphogenetic protein; Differentiation; Immortalized cell line; p53 gene; Vertebrata; Mammalia; TP53 Gene; Mouse; Animal; Rodentia; Bone; Cell differentiation; Metabolism; Cell line establishment; Tumor suppressor gene
• ISSN: 8756-3282; 1873-2763
• DOI: 10.1016/S8756-3282(97)00152-X

An immortalized cell line exhibiting a well-differentiated osteoblast-like phenotype was established from calvaria of
p53 tumor suppressor-deficient mice. This cell line, designated MMC2, showed several osteoblast-like properties such as high alkaline phosphatase activity, expression of type I collagen and osteocalcin mRNA, and differentiated in vitro to produce mineralized extracellular matrix. Alkaline phosphatase activity and the level of osteocalcin mRNA expression and the production of mineralized matrix were significantly enhanced by the addition of ascorbic acid. Although the cells proliferated rapidly and indefinitely, they did not grow in soft agar and were nontumorigenic in nude mice. These characteristics were equivalent to those observed in MC3T3-E1, a well-known osteoblast-like cell line. When inoculated in nude mice, however, MMC2 produced matured bone tissue, which was not observed in the case of MC3T3-E1. Expression of bone morphogenetic protein 2 and 4 and type IA receptor mRNA was demonstrated in cultured MMC2 cells. These results indicate that this new osteoblast-like cell line, MMC2, will be a unique material for the analysis of bone cell biology.