人脐血不同前体细胞体外诱导分化为树突状细胞的研究

Generation of dendritic cells derived from cord blood two different precursor

目的 从人脐血不同的前体细胞体外诱导分化树突状细胞 (DCs) ,并对生成的细胞进行鉴定 ,探讨脐血在DC体外大量扩增的组织来源作用。方法 用免疫磁珠法分离人脐血CD34+ 细胞 ,以GM CSF、TNF α、FL、SCF、IL 4诱导生成DC ;用Ficoll分离脐血单个核细胞 ,2h贴壁后的细胞 ,以GM CSF、IL 4、TNF α诱导生成DC。电镜观察细胞形态 ,流式细胞仪分析细胞表型 ,并检测其刺激同种T细胞增殖的能力。结果 人脐血CD34+ 细胞和贴壁细胞体外诱导生成的DC均具有典型的DC形态特征 ,表达高水平的DC特异性标志CDla ,MHCⅡ类抗原递呈分子HLA DR和CD80共刺激分子。两种方法获得的DC的形态、CDla表面抗原的表达没有显著的差异 ,两者均具有刺激同种T细胞增殖的能力 ,前者扩增DC的效率更高。结论 从人脐血中CD34+ 细胞和贴壁细胞两种DC的前体细胞均可诱生出DC 。

Objective To proliferate dendritic cells(DCs) from human cord blood two different precursor.Methods CD34 + cells were isolated from umbilical cord blood by using a high gradient magnetic cell sorting system (MACS) and cultured with cytokines including GM CSF,TNF α,FL,SCF and IL 4;cord blood mononuclear cells were cultured for 2 h and nonadheret cells were removed,and then cytokines including GM CSF,TNF α and IL 4 were added into the adheret cells for further culture.Cells harvested were identified for their properties by microscope,FACS,and mixed lymphocyte reaction(MLR).Resultes Dendritic cells induced from both cord blood CD34 + cells and adherent cells had typical morphological properties of DC,and highly expressed differential antigens.Nevertheless,the induced DCs had the capacity to stimulate proliferation of allogenetic T lymphocytes.The quantity of DCs induced from CD34 + cells cultured with cytokines including GM CSF,TNF α,FL,SCF and IL 4,were much more than that induced from adheret cells cultured with cytokines including GM CSF,TNF α and IL 4.Conclusion Two precursors in cord blood,CD34 + cells and adheret cells,can be induced to functional DCs,Human cord blood may be a good source of DCs.