Establishment and characterization of osteoblast-like cell lines from retrovirus (RFB MuLV)-induced osteomas in mice.
Cell lines were established by a two-step method from osteomas which had been induced by infection of mice with RFB MuLV, a bone-pathogenic, replication-competent murine retrovirus. The benign tumors, consisting of mature lamellar bone and surrounded by a thin periosteum, were cultured on sponges of denatured collagen type I fibres for up to 4 weeks. At this time osteoma cells had grown into the collagenous matrix. After release and further cultivation in monolayers, the cell lines established from these cultures varied in morphology; they expressed T1, collagen type I and type III, alkaline phosphatase, osteonectin and osteopontin mRNAs at variable levels, but not osteocalcin/BGP. They also showed alkaline phosphatase activity, but lacked responsiveness to parathyroid hormone. All cell lines established from infected mice expressed retroviral and c-myc mRNA and viral protein. In contrast to cells from control mice they showed an extended life span in culture. After growth in a three-dimensional (3-D) collagen sponge culture the cells formed an extracellular matrix containing collagen type I, alkaline phosphatase and osteocalcin/BGP. These data indicate that the two-step method facilitates the establishment of osteoblast-like cell lines from osteomas and calvaria of old mice, and provides means for further analyses of retrovirus-induced skeletal pathogenesis and bone induction.
Goralczyk
R
R
Institute of Molecular Virology, GSF-National Research Center for Environment and Health, Neuherberg, Germany.
Appold
A
A
Luz
A
A
Riemann
S
S
Strauss
P G
PG
Erfle
V
V
Schmidt
J
J
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001859:Bone Neoplasms / Q000473:pathology / Q000821:virology*; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D009052:Leukemia Virus, Murine / Q000502:physiology*; D051379:Mice; D008808:Mice, Inbred CBA; D008815:Mice, Inbred Strains; D010006:Osteoblasts / Q000166:cytology*; D010016:Osteoma / Q000473:pathology / Q000821:virology*; D010281:Parathyroid Hormone / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000096:biosynthesis; D012192:Retroviridae Infections / Q000821:virology*; D014407:Tumor Cells, Cultured; D014779:Virus Replication
D010281:Parathyroid Hormone; D016271:Proto-Oncogene Proteins c-myc; D000469:Alkaline Phosphatase