Human bone cell enzyme expression and cellular heterogeneity: Correlation of alkaline phosphatase enzyme activity with cell cycle

✍ Scribed by N. S. Fedarko; P. Bianco; U. Vetter; P. Gehron Robey

• Publisher: John Wiley and Sons
• Year: 1990
• Tongue: English
• Weight: 798 KB
• Volume: 144
• Category: Article
• ISSN: 0021-9541
• DOI: 10.1002/jcp.1041440115

✦ Synopsis

Alkaline phosphatase, long implicated in bioniineralization, is a feature of the osteoblast phenotype. Yet in cultured bone cells, only a fraction stain positive histochemically. To determine whether osteoblast enzyme expression reflects cellular heterogeneity with respect to cell cycle distribution or length of time in culture, the activities of a1 kaline phosphatase, tartrate-resistant and -sensitive acid phosphatases, and non-specific esterases were assayed kinetically and histochemically. In asynchronous subconfluent cultures, <15% of the cells stained positive and assayed activity was 0.04 l U i l 0 " cells/cm2. After 1 week, the percent of alkaline phosphatase positive-staining cells increased 5-fold, while activity increased 10-fold. Non-specific esterascs and tartrate-sensitive acid phosphatase were constitutive throughout time in culture, whereas tartrate-resistant acid phosphatase activity appeared after 2 weeks. Cell cycle analysis of human bone cells revealed a growth fraction of DO%, an S phase of 8.5 h, G , + 112 M of 4 h, and a G, of 25-30 h. In synchronous cultures induced by a Ihymidine-dphidicolin protocol, alkaline phosphatase activity dropped precipitously at M phase and returned during G,. A majority of the alkaline phosphatase activity lost irorn the cell surface at mitosis was recovered in the medium. Tartrate-sensitive acid phosphatase and non-specific esterase levels were relatively stable throughout the cell cycle, while tartrate-resistant acid phosphatase activity was not assayable at the density used in synchronous cultures. From these data, variations in alkaline phosphatase activity appear to reflect the distribution of cells throughout the cell cycle.