Modulation of cell phenotype in human osteoblast-like cells by the simian virus 40.
At present, the majority of in vitro research into bone metabolism is performed on either primary cultures of bone or osteosarcoma lines. A better model of the behaviour of normal bone cells would be a cell line derived from normal, adult bone that retained osteoblast-like characteristics. We infected a culture of bone cells from adult humans with simian virus 1613, a variant of the simian virus 40, and obtained 12 clones of variable morphology. The clones were maintained in culture for as long as 6 months. Population doubling times, synthesis of alkaline phosphatase and osteocalcin, secretion of mineral, morphology, and ability to withstand freezing were examined. SV/EC cell morphology varied from the polygonal, osteoblast-like to the bipolar, fibroblast-like. Population doubling times ranged from 0.55 to 2.8 days (compared with 3.9 days for the nontransformed human osteoblast-like cells). Synthesis of alkaline phosphatase varied but was less than that by the human osteoblast-like cells. With the exception of clone 11, all of the transformed clones synthesised mineral in vitro under mineralising conditions. Four clones showed increased synthesis of alkaline phosphatase and increased population doubling times after passaging. All of the clones were successfully frozen and thawed, but, unlike normal human osteoblast-like cells, none responded to stimulation with parathyroid hormone.(ABSTRACT TRUNCATED AT 250 WORDS)
Evans
C E
CE
Department of Orthopaedic Surgery, University of Manchester.
Ng
K
K
Allen
J
J
Gallimore
P
P
D000469:Alkaline Phosphatase / Q000096:biosynthesis; D002113:Calcification, Physiologic; D002455:Cell Division; D002470:Cell Survival; D002472:Cell Transformation, Viral; D002999:Clone Cells; D000242:Cyclic AMP / Q000378:metabolism; D005615:Freezing; D006801:Humans; D010006:Osteoblasts / Q000166:cytology / Q000502:physiology* / Q000821:virology; D015675:Osteocalcin / Q000096:biosynthesis; D010641:Phenotype; D013539:Simian virus 40 / Q000502:physiology*
D015675:Osteocalcin; D000242:Cyclic AMP; D000469:Alkaline Phosphatase