人CK8&18阳性胸腺上皮细胞对脐血CD34^+细胞向T细胞分化的影响  

The effects of the human CK8/18 positive thymic epithelial cells on differentiation of cord blood CD34^+ cells to T cells

作　　者：李倩如[1] 杜焱[2] 苏艳宾[1] 杜英[1] 张清勇[3] 李国喜 胡祥 

机构地区：[1]郑州大学基础医学院微生物学与免疫学教研室,郑州450001 [2]郑州大学第二附属医院放射科,郑州450004 [3]郑州大学第二附属医院胸外科,郑州450004 [4]深圳市北科细胞工程研究所,深圳518057

出　　处：《中国免疫学杂志》2011年第3期200-203,共4页Chinese Journal of Immunology

基　　金：国家自然科学基金项目(No.30570625);河南省自然科学基金(No.511042700);广东省自然科学基金(No.2007B031500015)资助

摘　　要：目的:通过CK8/18阳性胸腺上皮细胞与人脐血单个核细胞在Transwell板的共培养,探讨CK8/18阳性TEC对T细胞增殖分化的影响。方法:用胶原酶消化法分离纯化胸腺上皮细胞、免疫组化鉴定分离纯化的TEC,采用密度梯度离心法分离获得脐血单个核细胞,并采用免疫磁珠分选CD34+细胞,将TEC种植培养于Transwell双层培养板上层,使其均匀平铺于上层板底部,再将分选后的细胞加入上层小室,经过48小时的共培养,流式细胞术检测进入下室细胞的表型变化。结果:分离纯化的TEC经免疫组化鉴定CK8/18阳性。TEC与脐血单个核细胞共培养后,CD3+CD4-CD8-双阴性、CD3+CD4+CD8-单阳性细胞显著增加,CD3+CD4+CD8+双阳性细胞亚群细胞减少,CD8单阳性细胞增加不明显;CD45RA阳性细胞比率无显著变化,CD45RO阳性细胞比率显著增加。结论:CK8/18阳性TEC能选择促进脐血单个核细胞CD4+T细胞和CD45RO+细胞增殖。Objective：To study the effects of CK8/18 positive thymic epithlial cell（TEC） on T cells proliferation and differentiation,we established a model by co-cultured CK8/18 positive TEC and cord blood mononuclear cells（CBMC） in transwell co-culture system.Methods：Thymic epithelial cells were obtained through collagenase digestion culture and identified with TEC keratin（CK） and CK8/18 antigen.CD34＋ cells were collected and purified from CBMC by means of imunomagnetic beads on MiniMACS device.The original culture of CK8/18 positive TEC was put into transwell upper chamber（pore size 5 μm）,and then the CD34＋ cells were added to transwell upper chamber until the PC membrane were completely covered by TEC,when the cells into the lower chamber after 48 hours were detected with flow cytometry.Results：The TEC were anti-CK-positive and anti-CK8/18 staining positive.Compared with the control,the CD3＋CD4-CD8-double negative cells,CD3＋CD4＋CD8-single and CD45RO＋ cells that entered the transwell lower chamber increased significantly（P〈0.01） in transwell co-culture system.Conclusion：CK8/18 positive thymic epithelial cells can promote CBMC CD4＋ single-positive T cells and CD45RO＋ cells proliferation.

关 键 词：胸腺上皮细胞 脐血单个核细胞 T细胞亚群 

分 类 号：R392.3[医药卫生—免疫学]