华西口腔医学杂志

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口腔黏膜上皮细胞体外培养的影响因素分析及改进方法

王翔1, 高文信1, 王莉2, 魏秀峰1, 陈英新1, 王晓峰1   

  1. 1.吉林大学口腔医院黏膜病科, 吉林长春130041; 2.吉林大学基础医学院免疫学教研室, 吉林长春130021
  • 收稿日期:2007-02-25 修回日期:2007-02-25 出版日期:2007-02-20 发布日期:2007-02-20
  • 通讯作者: 高文信,Tel:0431- 8975882
  • 作者简介:王翔(1979-),男,新疆人,硕士研究生
  • 基金资助:

    国家“十五”科技攻关资助项目(2004BA720A28);吉林省科技发展计划资助项目(20050405- 1);吉林大学创新基金资助项目(419070200084)

Influential Factor Analysis and Improvement of Cultur al Method of Or al Mucosal Epithelial Cells

WANG Xiang1, GAO Wen- xin1, WANG Li2, WEI Xiu- feng1, CHEN Ying- xin1, WANG Xiao- feng1   

  1. 1. Dept. of Oral Medicine, School of Stomatology, Jilin University, Changchun 130041, China;2. Dept. of Immunology, School of Basic Medical Sciences, Jilin University, Changchun 130021, China
  • Received:2007-02-25 Revised:2007-02-25 Online:2007-02-20 Published:2007-02-20
  • Contact: GAO Wen- xin,Tel:0431- 8975882

摘要:

目的分析人口腔黏膜上皮细胞培养的影响因素,并建立稳定的细胞培养体系。方法新洁尔灭和硫酸庆大霉素联合处理口腔黏膜标本,比较不同浓度的Dispase对上皮的分离作用,酶消化法分离细胞,采用无血清培养液进行原代和传代培养。结果微生物污染得到有效控制,0.40%的Dispase的上皮分离效果优于0.25%的Dispase,细胞在短期内快速稳定增殖。结论人口腔黏膜上皮细胞的原代培养方法经改进后可简化操作,并提高成功率。

关键词: 细胞培养, 口腔黏膜, 上皮细胞

Abstract:

Objective To analyze influential factors of cultural method of human oral mucosal epithelial cells (hOMEC)and to establish a reliable cell culture system for hOMEC. Methods Benzalkonium bromide and gentamicin sulfate were used to prevent microbial contamination. Separations of epithelium from underlying connective tissues with Dispase at different concentration were compared. Enzyme digestion was used to isolate cells and ker - atinocyte- serum free medium(K- SFM)was employed for primary culture and subculture of hOMEC. Results Microbial contamination was under control. Separation of epithelium from underlying connective tissues with 0.40% Dispase was more complete than that of 0.25% Dispase. The cells grew fast and well in vitro. Conclusion The high successful culture of hOMEC and simplified procedures could be obtained with improvement of methods.

Key words: cell culture, oral mucosa, epithelial cell