中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (40): 7549-7553.doi: 10.3969/j.issn.1673-8225.2011.40.032

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

三维培养条件下胚胎鼠唾液腺上皮细胞的自组装

王军胜1,黄桂林2,张霓霓2,姚  礼2,唐岳鹏2   

  1. 1西安医学院附属医院口腔科,陕西省西安市710077
    2遵义医学院附属口腔医院口腔颌面外科,贵州省遵义市563003
  • 收稿日期:2011-02-16 修回日期:2011-04-19 出版日期:2011-10-01 发布日期:2011-10-01
  • 通讯作者: 黄桂林,博士,教授,遵义医学院附属口腔医院口腔颌面外科,贵州省遵义市 563003 chaojiehuanghgl@163.com
  • 作者简介:王军胜★,男,1980年生,甘肃省天水市人,汉族,2010年遵义医学院毕业,硕士,医师,主要从事口腔颌面部修复与重建方面的研究。 wswjs198028@163.com
  • 基金资助:

    贵州省优秀科技教育人才省长资金[黔省专合字(2008)113号];贵州省教育厅基金[(2006)354号]。

Self-organization and branching morphogenesis of primary salivary epithelial cells under three-dimensional culture

Wang Jun-sheng1, Huang Gui-lin2, Zhang Ni-ni2, Yao Li2, Tang Yue-peng2   

  1. 1Department of Stomatology, Affiliated Hospital of Xi'an Medical College, Xi'an  710077, Shaanxi Province, China
    2Department of Oral and Maxillofical Surgery, Stomatological Hospital, Zunyi Medical College, Zunyi  563003, Guizhou Province, China
  • Received:2011-02-16 Revised:2011-04-19 Online:2011-10-01 Published:2011-10-01
  • Contact: Huang Gui-lin, Doctor, Professor, Department of Oral and Maxillofical Surgery, Stomatological Hospital, Zunyi Medical College, Zunyi 563003, Guizhou Province, China chaojiehuanghgl@163.com
  • About author:Wang Jun-sheng★, Master, Physician, Department of Stomatology, Affiliated Hospital of Xi'an Medical College, Xi'an 710077, Shaanxi Province, China wswjs198028@163.com
  • Supported by:

    Guizhou Governor Funding for Outstanding Professionals in Education, No. (2008)113*; the Fund of Guizhou Education Bureau, No. (2006)354*

摘要:

背景:对唾液腺胚胎细胞在体外自组装及形态发生过程的理解,将有利于在体外构建唾液腺器官样组织的研究。
目的:探讨三维培养条件下SD大鼠胚胎鼠下颌下腺上皮细胞的自组装机制。
方法:将胚胎期15 d鼠的下颌下腺上皮细胞分为实验组(加入抗E-cadherin及抗β-catenin)和对照组(常规培养),在Matrigel中三维培养。
结果与结论:获得的胚胎鼠下颌下腺上皮细胞均表达CK19。实验组细胞未见分枝状及腺泡样结构出现;E-cadherin 和β-catenin荧光表达弱。对照组细胞在12 d出现上述结构;胞浆中E-cadherin、β-catenin荧光表达强表达。MOD值为实验组<对照组(P < 0.01)。提示E-cadherin,β-catenin在细胞的自组装过程中有重要作用。

关键词: 下颌下腺上皮细胞, 胚胎鼠, 三维培养, 自组装, E-cadherin, &beta, -catenin

Abstract:

BACKGROUND: To understand self-assembly and branching of primary salivary epithelial cells will provide insights into reconstructing artifical salivary gland orgnoid.
OBJECTIVE: To investigate the machanism of self-organization and branching mophogenesis of primary submandibular epithelial cells under three-dimensional condition.
METHODS: The embryonic day 15 (E 15) mouse submandibular gland epithelium were inoculated in matrigel for 3-dimantional culture with/without (Group A/Group B) anti-E-cadherin and and anti-β-catenin. The contructs of these cells aggregation were observed under inverted microscope at days 1, 6, 12. The contructs were observed with cytokeratin 19 immunohistochemistry staining, and identified with Cadherin and β-catenin immunostaining and confocal microscopy.
MAIN OUTCOME MEASURES: The constructs were observed under inverted microscope and analyzed by antibody of cytokeratin 19 with immunohistochemistry, and identified with cadherin and β-catenin immunostaining and confocal microscopy. In Group A, branching morphogenesis and acinar-like structure were not observed. In Group B, these contructs could be formed at day 12. In Group A, weak green fruorescence of E-cadherin and β-catenin expressed weakly. In Group B, they expressed strongly, and in the cytoplasm. MOD was lower in Group A than Group B (P < 0.01). E-cadherin and β-catenin are two important factors during the embryonic submandibular salivary gland epithelium self-organization.

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