Establishment and characterization of two immortalized cell lines of the osteoblastic lineage
| Autor: | A. Stutzer, Herbert Fleisch, H.L. Guenther, R. Friis, Willy Hofstetter, R. Schenk |
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| Rok vydání: | 1991 |
| Předmět: |
medicine.medical_specialty
Endocrinology Diabetes and Metabolism Cell Population Molecular Sequence Data Osteocalcin Genes myc Biology Dinoprostone Osteogenesis Internal medicine Bone cell medicine Animals Orthopedics and Sports Medicine education Cell Line Transformed education.field_of_study Osteoblasts Base Sequence Estradiol Osteoblast Alkaline Phosphatase Blotting Northern Molecular biology Immunohistochemistry Clone Cells Rats Blotting Southern Endocrinology medicine.anatomical_structure Cell culture Parathyroid Hormone biology.protein Alkaline phosphatase Collagen Immortalised cell line Adenylyl Cyclases |
| Zdroj: | Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research. 6(6) |
| ISSN: | 0884-0431 |
| Popis: | Osteoblastic cells were cloned by culturing rat calvariae cells in agarose in the presence of TGF-beta and EGF. Two bone cell lines were established by immortalizing such an osteoblastic clonal cell population by the introduction of the avian v-mycOK10 gene in the form of a mouse ecotropic retrovirus. Although originating from the same clonal cell population, the two lines exhibited somewhat differing properties. IRC10/30-myc1 expressed alkaline phosphatase (AP), showed PTH- and PGE2-induced cAMP production, synthesized mainly collagen type I and a minor fraction of type III, and produced mRNA for the bone-specific protein osteocalcin. IRC10/30-myc3 did not express AP, showed no PTH responsiveness, and synthesized only about one-third as much collagen as IRC10/30-myc1 (4 versus 12% of total protein synthesis). However, the cell line IRC10/30-myc3 was induced to synthesize cAMP by PGE2 and produced osteocalcin mRNA. When cultured in vivo in diffusion chambers, both lines proved to be osteogenic. Besides bone, both lines also formed cartilage and fibrous tissue. Thus, by immortalizing a clonal cell population of the osteoblastic phenotype, cell lines expressing varying properties can emerge. Furthermore, the expression of alkaline phosphatase and PTH-inducible adenylate cyclase are not prerequisites for a cell to form bone in vivo. Finally, cells expressing the phenotype of differentiated osteoblasts, including osteocalcin synthesis, still have a multipotential differentiation capacity and form bone and cartilage in vivo. |
| Databáze: | OpenAIRE |
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