Characteristics and culture of osteoblasts derived from avian long bone

A method is presented for isolating primary osteoblasts from the periosteal surface of chick tibia. The culture system identified supports both cell proliferation and phenotype retention. Cell numbers increased 8-fold in Week 1 and 20-fold over a total of 12 days. Well-established osteoblast markers, alkaline phosphatase staining, gamma-carboxyglutamic acid, osteocalcin, type I collagen, and parathyroid hormone binding were detected. Osteocalcin, gamma-carboxyglutamic acid, and type I collagen were present on culture Day 4, and were increased in amount by Day 8, but were similar to the earlier level on Day 12, suggesting that the phenotype may revert to a less differentiated state by 12 days in culture. Alkaline phosphatase staining was intense at all three assay times, however. During the last 4 days of the 12-day culture period, proliferation rates were higher than in the previous 8 days.