Enhancement of in vitro osteoblastic potential after selective sorting of osteoblasts with high alkaline phosphatase...

In this article we describe the expansion of in vitro osteogenic capability of human osteoblasts (HOS cells) after sorting by fluorescence-activated cell sorting (FACS) with the osteoblastic marker of human bone alkaline phosphatase (AP) monoclonal antibody. After culturing for 7 days, the HOS cells were incubated with fluorescein isothiocyanate (FITC)-labeled AP monoclonal antibody. The antibody recognized the cells with high AP activity (high AP cells), which were about 76% of the total cells. After the HOS cells were sorted, the high AP cells could be recovered, and almost all of them reacted strongly with the AP antibody. Therefore, we were able to condense the high AP cells about 1.3 times. We further cultured the sorted cells as well as the unsorted control cells. After the initial seeding, the culturing periods for both groups of cells were 20 days. At the end of this period, we measured AP activity per DNA and osteocalcin contents. In contrast to the low condensation ratio of the high AP cells in the sorted fraction, the AP activity and osteocalcin contents were about nine times and four times greater than those of the unsorted cells, respectively. These results demonstrated that using the sorting technique to isolate the high AP cells might be a useful method for applications in bone tissue engineering.