Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (32): 5993-5998.doi: 10.3969/j.issn.2095-4344.2012.32.020

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Primary culture of human periodontal ligament stem cells and dental pulp stem cells by tissue enzymatic digestion method

Li Ying1, Gu Zi-ya2, Lü Qiu-feng2, You Jin-biao1, Liu Fang-fei3   

  1. 1Institute of Biological Engineering, Zhejiang Academy of Medical Sciences, Hangzhou 310012, Zhejiang Province, China;
    2Hangzhou Stomatological Hospital of Zhejiang Province, Hangzhou 310012, Zhejiang Province, China;
    3Tongde Hospital of Zhejiang Province, Hangzhou 310012, Zhejiang Province, China
  • Received:2011-12-08 Revised:2012-01-06 Online:2012-08-05 Published:2012-08-05
  • About author:Li Ying★, Master, Institute of Biological Engineering, Zhejiang Academy of Medical Sciences, Hangzhou 310012, Zhejiang Province, China Liying19831121@yahoo.com.cn

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Abstract:

BACKGROUND: Tissue enzymatic digestion is an efficient primary culture method. This method can be used to elevate the success rate of primary cultured periodontal ligament stem cells and dental pulp stem cells.
OBJECTIVE: To compare the success rate of primary culture, morphological properties, cell growth between periodontal ligament stem cells and dental pulp stem cells, and identify the two isolated cell lines with different cell markers.
METHODS: Totally 20 health teeth were obtained by tooth extraction for orthodontic purposes. Among them, 16 teeth were pooled for periodontal ligament tissue, and four were pooled for dental pulp tissue. Periodontal ligament stem cells and dental pulp stem cells were isolated from these tissues by enzymatic digestion. Cell growth curve was obtained by the MTT method. Different cell markers of periodontal ligament stem cells and dental pulp stem cells were characterized by RT-PCR, immuocytochemistry staining and immunofluorescence staining.
RESULTS AND CONCLUSION: The success rate of the primary cultured dental pulp stem cells was significantly higher than that of the periodontal ligament stem cells, and the dental pulp stem cells showed a stronger capability of proliferation than the periodontal ligament stem cells. Purified periodontal ligament stem cells and dental pulp stem cells both demonstrated normal morphological and biological characteristics, and expressed cell markers CD105, CD73, vimitin and Stro-1, but did not express CK-17 and CD45. It is indicated that tissue planting after tissue enzymatic digestion is an efficient method for primary culture of periodontal ligament stem cells and dental pulp stem cells.

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