West China Journal of Stomatology

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Culturing and characterization of human periodontal ligament stem cells and investigating their chemotactic responses to bone morphogenetic protein-2

Du Lingqian1,2, Yang Pishan1,3, Zhao Ning1,3, Ge Shaohua1,3   

  1. 1. Dept. of Periodontology, School of Stomatology, Shandong University, Jinan 250012, China; 2. Dept. of Periodontology, The  Second Hospital of Shandong University, Jinan 250012, China; 3. Shandong Provincial Key Laboratory of Oral Biomedicine, Jinan 250012, China
  • Received:2012-02-25 Revised:2012-02-25 Online:2012-02-01 Published:2012-02-01
  • Contact: Ge Shaohua,Tel:0531-88382368
  • About author:Du Lingqian(1987—),女,山东人,硕士

Abstract:

Objective To investigate the chemotactic response of human periodontal ligament stem cells(PDLSCs)to bone morphogenetic protein-2(BMP-2). Methods Human PDLSCs were obtained from clinically healthy premolars extracted for orthodontic reasons and used to isolate PDLSCs by limited dilution method. The expression of Vimentin and stem cell marker STRO-1 on PDLSCs were demonstrated with immunocytochemical staining. Differentiation assay was used to detect the differentiation potential of PDLSCs. Cloning formation experiment and 5-bromo-2-deoxyuridine (BrdU) incorporation assay were used to determine the stem cell characteristics of PDLSCs. The chemotactic effect of BMP-2 on PDLSCs was detected by using a 24-multiwell Transwell cell culture chamber. The number of net migrated cells was counted in different microscope fields. Results Human PDLSCs displayed positive staining for Vimentin and expressed the stem cell marker STRO-1. These cells differentiated into osteoblasts and adipocytes under defined culture conditions, possessed high self-renewal potential and formed single-cell colonies in vitro. The number of cells migrating at concentrations of 100, 200 ng·mL-1 of BMP-2 in Transwell cell culture chamber was significantly higher than that of negative control (P<0.01). Conclusion BMP-2 may participate in regulating chemotaxis of human PDLSCs.

Key words: bone morphogenetic protein-2, periodontal ligament stem cells, chemotaxis