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Enhancement of In Vitro Osteoblastic Potential After Selective Sorting of Osteoblasts With High Alkaline Phosphatase Activity From Human Osteoblast-Like Cells

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Abstract:

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.

Keywords: Alkaline phosphatase; Cell sorting; Flow cytometry; Osteoblast; Tissue engineering

Document Type: Research Article

DOI: http://dx.doi.org/10.3727/000000004783983756

Affiliations: Tissue Engineering Research Center (TERC), National Institute of Advanced Industrial Science and Technology (AIST), Nakoji 3-11-46, Amagasaki, Hyogo 661-0974, Japan

Publication date: January 1, 2004

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  • Cell Transplantation publishes original, peer-reviewed research and review articles on the subject of cell transplantation and its application to human diseases. To ensure high-quality contributions from all areas of transplantation, separate section editors and editorial boards have been established. Articles deal with a wide range of topics including physiological, medical, preclinical, tissue engineering, and device-oriented aspects of transplantation of nervous system, endocrine, growth factor-secreting, bone marrow, epithelial, endothelial, and genetically engineered cells, among others. Basic clinical studies and immunological research papers are also featured. To provide complete coverage of this revolutionary field, Cell Transplantation will report on relevant technological advances, and ethical and regulatory considerations of cell transplants. Cell Transplantation is now an Open Access journal starting with volume 18 in 2009, and therefore there will be an inexpensive publication charge, which is dependent on the number of pages, in addition to the charge for color figures. This will allow work to be disseminated to a wider audience and also entitle the corresponding author to a free PDF, as well as prepublication of an unedited version of the manuscript.

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