中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (32): 5993-5998.doi: 10.3969/j.issn.2095-4344.2012.32.020

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

酶解组织块法原代培养人牙周膜干细胞和牙髓干细胞

李 颖1，谷子芽2，吕秋峰2，尤金彪1，刘芳菲3   

  1. 1浙江省医学科学院生物工程所，浙江省杭州市 310012；
    2浙江省杭州口腔医院， 浙江省杭州市 310012；
    3浙江省立同德医院，浙江
  • 收稿日期:2011-12-08 修回日期:2012-01-06 出版日期:2012-08-05 发布日期:2012-08-05
  • 作者简介:李颖★，女，1983年生，浙江省湖州市人，汉族，2008年浙江大学毕业，硕士，研究实习员，主要从事干细胞研究。 Liying19831121@yahoo.com.cn

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

PDF

706

被引次数

21

摘要：

背景：酶解组织法是一种高效的原代细胞培养方法，利用该法可提高牙周膜干细胞和牙髓干细胞的培养成功率。
目的：比较牙周膜干细胞和牙髓干细胞的原代培养成功率、细胞形态、生长特征，并对其进行多种细胞标志的鉴定。
方法：取因正畸需要拔除健康牙20颗，其中16颗用于牙周膜干细胞原代培养，4颗用于牙髓干细胞培养。酶解组织块法培养两种细胞，MTT法检测两种细胞的生长曲线，利用RT-PCR法、细胞免疫组化法、细胞免疫荧光法鉴定两种细胞的标志。
结果与结论：牙髓干细胞的原代培养成功率明显高于牙周膜干细胞，显示更快的生长能力。通过酶解组织法分离纯化的两种干细胞均显示正常的细胞形态和生长特征，表达相应的细胞标志CD105，CD73，波形丝蛋白vimitin，Stro-1，不表达角蛋白CK-17和CD45。提示酶解组织块法是一种高效的原代培养牙周膜干细胞和牙髓干细胞的方法。

关键词: 牙周膜干细胞, 牙髓干细胞, 酶解组织块法, 原代培养, 细胞形态, 干细胞

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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