Biochemical characterization of embryonic chick calvarial cells

Yu, Jae Hyung; Kim, Jung Kun; Cha, Kyung Suk

Abstract

Chicken calvarial bone is known to contain various cell types, but their exact composition is unknown. By characterizing the chicken calvarial bone biochemically, it can be used to study biochemical, histochemical actions of bone cells in general. Calvaria of I8-day-old white leg horn embryo was aseptically dissected and bone cell populations were isolated by sequential enzymatic digestion. Histochemical study for osteoclast-like bone cell population was performed with tartrate resistant acid phosphatase(TRAP) stain and for osteoblast-like bone cell population, alkaline phosphatase(ALP) stain was performed. Biochemical study for osteoblast-like bone cell population was performed using alkaline phosphatasetal.P) assay. Following conclusions were obtained from this study. 1. TRAP positive multi and mononuclear cells were mostly observed in group I and II, indicating that osteoclast-like bone cell population is mostly found in these groups. 2. All the cultured groups showed almost equal ALP activities and were positive for ALP stain, indicating that osteoblast-like bone cell population is evenly dispersed in all culture groups. 3. Experimental group treated with I,25(OH)2D3 showed increase in ALP activity in contrast to the control group, confirming previous studies that I,25(OH)2D3 increases ALP activities in vitro bone cultures. 4. Results from von Kassa's stain indicated that in vitro bone formation had occured after 3 weeks of culture with beta glycero phosphate.