颅神经嵴干细胞的体外培养及其多向分化潜能研究

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。

In vitro Culture and Biological Characteristics of Cranial Neural Crest Stem Cell

Objective According to development biology, cranial neural crest stem cell (CNCSC) can differentiate into precursor cells of tooth, jaw and peripheral nerve system, but in vitro study is less reported. In the present study, CNCSC were dissociated and cultured in vitro,and the biological characteristics of CNCSC was investigated.Methods Cranial neural tubes, dissected from mouse E9d, were explanted onto fibronectin-coated dishes. CNCSCs emigrated from the explanted neural tubes, cultured 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 differentiation of CNCSC in vitro, especially providing a cell source for investigating tooth/jaw regeneration.