摘要
目的体外培养近交系大鼠胚胎腹侧中脑前体细胞(VMP)并诱导其分化为多巴胺能神经元(DN),为研究DN定向分化的分子机制提供细胞模型。方法取材胎龄11d的近交系大鼠胚胎VMP,体外用碱性成纤维细胞生长因子(bFGF)增殖培养7d后换用L-抗坏血酸-2-磷酸酯倍半镁盐(AA-2P)诱导分化为DN,随后进行免疫荧光染色鉴定。结果细胞总数扩增49.76倍,免疫荧光染色显示β-TubulinⅢ阳性的神经元中(71.33±20.42)%为TH阳性的DN,后者占细胞总数的(24.85±12.85)%。结论近交系大鼠VMP经体外原代培养能够得到较高比例的DN,可作为深入研究DN定向分化分子机制的细胞模型。
Objective Toestablish a cell model for studying the molecular mechanism of committed differentiation of dopaminergic neurons（DNs）, by means of in vitro culturing ventral mesencephalic precursors （VMPs） from inbred rat embryos. Methods VMPs harvested from ventral mesencephalon of El I inbred rat embryos were proliferatively cultured in vitro with bFGF for 7 days, and differentiated into dopaminergic neurons in free-bFGF medium supplemented with AA-2P. Immunofluorescence staining was performed to evaluate the yield of DNs. Results The total cell number increased about 49.76 folds. Among these cells, nearly （24.85±12.85）% were tyrosine hydroxylase （TH）-positive cells （DN）, which made up about （71.33±20.42）% of β-Tubulin Ⅲ-positive neurons. Conclusion VMPs of inbred rat embryos can differentiate into considerable DNs in vitro, which therefore can be used as an ideal cell model for exploring the molecular mechanism of DN differentiation.
出处
《中国微侵袭神经外科杂志》
CAS
2006年第7期321-323,共3页
Chinese Journal of Minimally Invasive Neurosurgery
基金
江苏省自然科学基金资助项目(BK2004043)
