Approach to get mineralized tissue using pulp cells

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The purpose of this study was to get pluripotent cells from human pulp tissues by simple technique. To get the mils, we separated cells by the indication for cell surface antigens. Using Fluorescence Activated Cell Sorter (FACS) and Magnetic Cell Sorting (MACS) systems, we have done the following experiments(1) We collected human pulp tissues aseptically, allow for them by outgrowth, continue to growth, then analysis them by FACS. We used CD73, 74, 106, 146, 166, 271, and STRO-1 antigen. As the results, the expression level of six antigen except CD166 were quite low, however positive expression were detected by 20% of CD166 stained cells.(2) We conjugated the cells to the above antigen except CD73, and apply them to MACS system. We continued to growth the antigen positive cells, and differentiated them to osteoblasts. As the results, Alizarin red staining levels were lower in CD74 and 106 positive cells than in CD74 and 106 negative cells. CD146 and STRO-1 positive cells show same Alizarin red staining levels compared with CD146 and STRO-1 negative cells. CD166 and 271 positive cells show obvious strong Alizarin red staining compared with CD166 and 271 negative cells. These CD166 and 271 positive cells show high activity of Alkaline phosphates. Alizarin red staining of CD166 and 271 positive cells were obviously higher than non selected pulp cells, especially in CD166 positive cells.Taken together from these results show that MACS system utilized for CD166 is a useful method to sort out high capacity of osteogenic cells from human pulp.