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树突状细胞体外定向诱导扩增及其分离纯化 |
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白慈贤 冯凯 李梁 裴雪涛
摘 要 目的：探讨定向诱导及纯化树突状细胞(DC)的方法，为进一步研究树突状细胞的功能、特性及临床应用提供技术方法。方法：利用免疫磁珠法(MACS)分离纯化脐血CD34+细胞，在液体培养体系中加入FL、GM-CSF、TNF-α、IL-4及SCF，培养12 d后光镜检测细胞形态及流式细胞仪分析表面标志(CDla、HLA-DR及CD80)；利用抗树突状细胞单克隆抗体(X-11)及免疫磁珠分离纯化树突状细胞，流式细胞仪检测纯化后DC的纯度。结果：经体外定向诱导扩增，可成功地将CD34+细胞定向诱导为树突状细胞，CDla+细胞的比例可升至(27.18±1.56)%〔对照为(0.65±0.38)%)〕，诱导出的DC具有典型的树枝状突起，有较高的CDla、HLA-DR及CD80表达，经免疫磁珠分选，树突状细胞(CDla+)的纯度可达(94.61±2.24)%以上。结论：经特定细胞因子的组合，可将CD34+细胞定向诱导成DC，并可通过免疫磁珠法分选出纯度达90%以上的DC。
关键词 树突状细胞 诱导分化 分离纯化
中国图书分类号 R392-33
Studies on the committed differentiation and purification of dendritic cells
BAI Ci-Xian，FENG Kai，LI Liang et al.
Beijing Institute of Radiation Medicine，Beijing 100850
Abstract Objective:To elucidate and provide methods for inducing CD34+ cells to differentiate to dendritic cells(DCs) and for purifying DC.Methods:CD34+ cells were isolated from umbilical cord blood by using a high-gradient magnetic cell sorting system(MACS) and cultured with cytokines including FL、GM-CSF、TNF-α、IL-4 and SCF in a liquid culture system.Cells harvested after 12 days were detected for morphological and phenotypic analysis by microscope，and FACS respectively.DCs were purified by using monoclonal antibody X-11 and magnetic beads.Results:CD34+ cells can be induced into DCs in the culture system and the DCs can be purified at the grade of (94.61±2.24)% by using magnetic beads labeled monoclonal antibody mediated cell sorting system.The induced cells have the typic morphology and high expression of CDla，HLA-DR and CD80.Conclusion:It is possible to induce CD34+ cells to DCs and induced DCs can be purified at the grade of about 90%.
Key words Dendritic cells Induction of differentiation Purifying
树突状细胞(Dendritic cells，DC)是目前为止发现的功能最强的专职抗原提呈细胞(Antigen presenting cell，APC)，它对诱导初次免疫应答具有独特的功能，在对抗原的摄取、加工、处理及提呈过程中，DC既具有一般APC的共性，也表现出一些独特的生物学特性。由于它在机体的抗感染免疫及抗肿瘤免疫应答中的作用极其重要而[1] [2] [3] 下一页
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