华西口腔医学杂志

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颅神经嵴干细胞的体外培养及其多向分化潜能研究

田卫东,江宏兵,刘磊,汤炜,李晓东   

  1. 口腔生物医学工程教育部重点实验室,四川大学,四川成都610041
  • 收稿日期:2004-06-25 修回日期:2004-06-25 出版日期:2004-06-20 发布日期:2004-06-20
  • 通讯作者: 田卫东,Tel:028-85503406
  • 作者简介:田卫东(1963-),男,云南人,教授,博士
  • 基金资助:

    国家科技部重大基础研究前期专项资金资助项目 (2002CCC00700)

In vitroCulture and Biological Characteristics of Cranial Neural Crest Stem Cell

TIANWei-dong,JIANGHong-bing, LIU Lei, TANG Wei, LI Xiao-dong   

  1. Key. Laboratory of Oral Biomedical Engineering Ministry ofEducation, Sichuan University, Chengdu610041, China
  • Received:2004-06-25 Revised:2004-06-25 Online:2004-06-20 Published:2004-06-20

摘要:

目的 分离培养颅神经嵴干细胞,探讨其体外生物学特征,为进一步研究其牙、颌向分化提供细胞来源。方法 取小鼠胚胎颅段神经管,组织块法无血清条件培养获颅神经嵴干细胞,形态学、BurdU掺入及免疫细胞化学法鉴定其生物学性状。结果 原代培养的颅神经嵴干细胞为梭形成纤维样细胞,表现为克隆样生长、自我更新能力和多向分化能力等干细胞性状,且特异性标记物HNK-1呈阳性表达。结论 本研究成功地获取保持有多潜能干细胞性状的颅神经嵴细胞,为深入研究其牙、颌向分化奠定基础。

关键词: 颅神经嵴, 干细胞, 生物学特征, 分化

Abstract:

Objective According to development biology, cranial neural crest stem cell (CNCSC) can differentiate into pre- cursor cells of tooth, jaw and peripheral nerve system, butin vitrostudy is less reported. In the present study, CNCSCwere dis- sociated and culturedin vitro,and the biological characteristics of CNCSCwas investigated.Methods Cranial neural tubes, dis- sected from mouse E9d, were explanted onto fibronectin-coated dishes. CNCSCs emigrated from the explanted neural tubes, cul- tured in a serum-free medium containing modified DMEM/F12.Biological characteristics of CNCSC were detected by morphology, nuclear labeled with BurdU and immunocytochemistry.Results Fibroblast-like CNCSCs demonstrated the characteristics of stem cell, such as clonality, self-renewal and multipotentiality. The result of immunocytochemical stain showed that CNCSC expressed HNK-1 antigen.Conclusion CNCSC were cultured successfully, providing a experimental basis for study on tooth/jaw-like dif- ferentiation of CNCSCin vitro, especially providing a cell source for investigating tooth/jaw regeneration.

Key words: cranial neural crest, stem cell, biological characteristics, differentiation