Cv. Gay et al., CHARACTERISTICS AND CULTURE OF OSTEOBLASTS DERIVED FROM AVIAN LONG-BONE, In vitro cellular & developmental biology. Animal, 30A(6), 1994, pp. 379-383
A method is presented for isolating primary osteoblasts from the perio
steal surface of chick tibia. The culture system identified supports b
oth cell proliferation and phenotype retention. Cell numbers increased
8-fold in Week 1 and 20-fold over a total of 12 days. Well-establishe
d osteoblast markers, alkaline phosphatase staining, gamma-carboxyglut
amic acid, osteocalcin, type I collagen, and parathyroid hormone bindi
ng were detected. Osteocalcin, gamma-carboxyglutamic acid, and type I
collagen were present on culture Day 4, and were increased in amount b
y Day 8, but were similar to the earlier level on Day 12, suggesting t
hat the phenotype may revert to a less differentiated state by 12 days
in culture. Alkaline phosphatase staining was intense at all three as
say times, however. During the last 4 days of the 12-day culture perio
d, proliferation rates were higher than in the previous 8 days.