鸡胚颅盖骨成骨细胞样细胞培养液的生化分析

Biochemical Analysis of the Extracellular-matrix of Isolated Osteoblastic Cells in Cultured Calvarium of Chicken Embryos in Vitro

计数１０＾６鸡胚颅盖骨成骨细胞样细胞于培养瓶内培养，共１０瓶。每周测培养液中透明质酸，羟脯氨酸的含量和碱性磷酸酶的活力。结果；培养液的透明质酸含量从第２周的１３９．５２ｎｇｆ／ｍｌ上升至第９周的２７１１．９２ｎｇ／ｍｌ、表面细胞培养时，透明质酸释放量培养时间的延长而增加。

The isolated osteoblastic cells from embryonic chicken calvaria bone were cultured in vitro. The medium was changed twice every week. The concentrations of hyaluronic acid and hydroxyproline and the activity of alkaline phosphatase in the extracellular-matrix were determined once a week. At day 14, hyaluronic acid was detected in the medium at a concentration of 139. 52ng/ml and the concentration was increased to 2711. 9ng/ml after 9 weeks in culture. This result indicates that when the cells were cultured in vitro, hyaluronic acid was released into the extracellular-matrix and the amounts kept increasing. From day 14 to 9 weeks, hydroxyproline concentration in the medium maintained at 1μg/ml. This suggests that the amounts of hydroxyproline secreted into the medium were not related to the length of the process. After 2 weeks in culture, theactivity of alkaline phosphatase was relatively low, then it was increased gradually and reached maximal value at day 28. Thereafter, this value at this level fluctuated until 9 weeks in culture. These data show that alkaline phosphatase is related to the activity of bone formation.