Isolation And Identification Of Periodontal Ligament Stem Cells

Objective:Periodontal ligament cell populations were isolated from the periodontal membrane of extracted teeth and were purified; On this basis, the purified cells were identified. This paper confirmed that the cells were "stem cells" cells, to provide a basis for follow-up study of periodontal tissue engineering experimental.Methods:1. Separation and purification of human periodontal ligament stem cells Collecting the permanent teeth removal, without caries and periodontal disease, was indicated in the clinical cases due to orthodontic or impacted. Take the periodontal membrane on the middle 1/3 of the root surface. We separated cells within the periodontal tissue membrane by enzymatic, purified cells by limiting dilution cloning. The growth of periodontal ligament cell populations and purified periodontal ligament cells were analyzed by MTT.2. preliminary identification of Periodontal ligament stem cells:Cell cycle and cell surface markers were detected by flow cytometry. Osteogenic medium induced cells to identified cells.Results:1. Cells purified by limiting dilution cloning appeared Spindle. The results show that the cell is a slow proliferation cycle stem cells by MTT detection.2. The cells purified were detected cell surface markers and cell cycle characteristics by flow cytometry. The result demonstration that the cells accordance with characteristics of stem cell.3. Purified cells were induced by osteogenic medium, cells were unipolar, there have been needle the size of calcium nodules, confirmed that the purified cells possess differentiation potential.Conclusion:1. Cells which maintain their biological characteristics were effectively isolated by Enzymatic and purified by limiting dilution cloning. And the cells have characteristics of stem cell cycle and potential of Osteogenic differentiation.2. Cultured periodontal ligament stem cells provide the basis for tissue engineering.