中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (45): 8464-8468.doi: 10.3969/j.issn.1673-8225.2010.45.024

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

人下颌下腺干/祖细胞的分离及培养

王春宇1，2，黄桂林1，张霓霓1，张跃荣1，宋庆高1，潘光华1，方  宁3   

  1. 1遵义医学院附属口腔医院口腔颌面外科，贵州省遵义市   563003；2遵义市口腔医院口腔颌面外科，贵州省遵义市     563000；3遵义医学院贵州省细胞工程重点实验室，贵州省遵义市     563003
  • 出版日期:2010-11-05 发布日期:2010-11-05
  • 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: 王春宇★，男，1973年生，贵州省遵义市人，汉族，2009年遵义医学院毕业，硕士，医师，主要从事唾液腺组织工程方面的研究。 chun5873@126.com
  • 基金资助:

    2009年贵州省教育厅自然科学类科研项目，项目批准号(20090112)， 课题名称：唾液腺干/祖细胞的自组装和形态发生及分化研究。

Isolation and culture of human submandibular gland stem/progenitor cells

Wang Chun-yu 1, 2, Huang Gui-lin1, Zhang Ni-ni1, Zhang Yue-rong1, Song Qing-gao1, Pan Guang-hua1, Fang Ning3   

  1. 1 Department of Oral and Maxillofacial Surgery, Stomatological Hospital, Zunyi Medical College, Zunyi  563003, Guizhou Province, China; 2 Department of Oral and Maxillofacial Surgery, Zunyi Municipal Stomatological Hospital, Zunyi  563000, Guizhou Province, China; 3 Guizhou Provincial Key Laboratory of Cell Engineering, Zunyi Medical College, Zunyi  563003, Guizhou Province, China
  • Online:2010-11-05 Published:2010-11-05
  • Contact: Huang Gui-lin, Doctor, Professor, Department of Oral and Maxillofacial Surgery, Stomatological Hospital, Zunyi Medical College, Zunyi 563003, Guizhou Province, China chaojiehuanghgl@163.com
  • About author:Wang Chun-yu★, Master, Physician, Department of Oral and Maxillofacial Surgery, Stomatological Hospital, Zunyi Medical College, Zunyi 563003, Guizhou Province, China; Department of Oral and Maxillofacial Surgery, Zunyi Municipal Stomatological Hospital, Zunyi 563000, Guizhou Province, China chun5873@126.com
  • Supported by:

     the Natural Science Research Program of Guizhou Province in 2009, No. 20090112*

PDF

492

被引次数

12

摘要：

背景：近年来国内外学者研究证实了唾液腺组织中的确存在一类具有成体干细胞特性的唾液腺细胞，但是这类干细胞的研究多集中在动物的唾液腺，而人类唾液腺干细胞的生物学特征研究较少。
目的：体外分离、培养人唾液腺干/祖细胞，并观察其生物学特征。
方法：从流产胎儿下颌下腺中分离、单克隆培养获得人唾液腺干/祖细胞，应用免疫荧光染色方法检测上皮细胞标志CK19，Laminin的表达，流式细胞术检测CD29，CD90.1和 CD117的表达。绘制人唾液腺干/祖细胞生长曲线，评价体外增殖能力。
结果与结论：人唾液腺干/祖细胞胞浆表达上皮细胞标志CK19及Laminin；流式细胞术结果显示，CD90.1，CD29及CD117的表达率分别为99.69%，98.83%及67.27%；体外培养也表现出高增殖能力、克隆形成能力和较高的传代数等组织干细胞特性。提示人胎儿下颌下腺中能分离出具有组织干细胞特性的细胞。

关键词: 胎儿下颌下腺, 人唾液腺干/祖细胞, 分离培养, 体外, 生物学特征

Abstract:

BACKGROUND: Recently, scholars have verified that salivary gland cells that have adult stem cell characteristics exist in salivary gland tissues. However, these stem cells mainly focus in animal salivary gland. There are a few studies concerning biological characteristics of human salivary gland stem cells.
OBJECTIVE: To isolate and culture human salivary gland stem/progenitor cells in vitro, and to investigate the biological characteristics of these cells.
METHODS: Human salivary gland stem/progenitor cells were harvested from submendubular glands of the legally abortion fetus for monoclonal culture. Epithelial cell marker cytokeratin 19 (CK19) and Laminin were analyzed by immunohistochemistry. Surface markers CD29, CD90.1 and CD117 expressions were detected by flow cytometric analysis. Growth curve of human salivary gland stem/progenitor cells was drawn, and proliferation ability in vitro was evaluated.
RESULTS AND CONCLUSION: The human salivary gland stem/progenitor cells expressed intracellular Laminin and CK19. Flow cytometric analysis results revealed that the percentage of cells positive for CD90.1, CD29 and CD117 were 99.69%, 98.83% and 67.27%. The cells had perfect ability of proliferation, clonality and passage in vitro. These indicate that these cells from fetus submandibular glands have the characteristics of tissue stem cells.

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