Mineralization and metabolic response in serially passaged adult rat bone cells
Cell populations derived from adult rat bone were grown in cell culture and characterized with respect to their morphology and response to hormones. The cells were isolated from adult rat calvaria by mechanical rather than enzymatic methods. Cultures were initiated in modified BGJb medium supplemented with fetal bovine serum. These cultures and several cloned populations derived from them retained the ability to mineralize in vitro even after extended serial passage.
Cultures derived from an osteoblast-enriched population showed an initial positive cAMP response to PTH and PGE2, but not to TCT. The PTH and PGE2 responses diminished with serial passage. The PTH response was no longer measurable at passage 6, and the PGE2 response was not evident in passage 11. In one clone, the PGE2 response persisted through passage 16. Adult rat skin fibroblasts cultured similarly did not respond to PTH or TCT, but still had a significant PGE2 response through passage 21.
The cultured cells formed multiple layers with localized areas of higher cell density. Mineral plaques with major diameters as great as 0.75 mm were evident in the areas of greater cell density. Less extensive mineral deposits were present throughout the culture. The mineral plaques consisted of apatite-like crystals deposited on an organic matrix. Matrix vesicles and mineralized spherules appeared to be associated with initial mineral deposition. The spherules apparently coalesced to form more complex mineralized structures. A limited amount of mineralization also was observed in rat skin fibroblast cultures.
Key wordsMineralization Bone culture Osteoblast Matrix vesicles Hormone response
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