临床级树突状细胞的分离和培养.pdfVIP

# 458 # 2004 31 8 ## 1, 2 2* 1, 2 2 1, 2 2 王 坤 , 吴一龙 , 周 清 , 林嘉颖 , 徐崇锐 , 杨学宁 Large-scale Generation of Mature Clinical Grade Dendritic Cells 1, 2 2 1, 2 2 1, 2 2 WANG Kun , WU Y-i l ng , ZHOU Qing , LIN Jia-ying , XU Ch ng-rui , YA NG Xue-ning 1. 3 rd H osp ital af f iliated of S un Yat-sen University , Guangz hou 5 10630, China; 2. L ung Cancer Research nstitute Cancer Center, Guang dong Prov incial Peop le. s H osp ital ( * Cor resp ond author ) Abstract: Objective T study the meth d f r generati n large-scale mature dendritic cells fr m n npr lifera- ti n pr genit rs in patient. s bl d. Methods The pr cedure uses 5% AB human plasma in the place f 10% fetal calf serum and inv lves tw steps. The first step is t w rk n a 5~ 7-day-culture f plastic-adherent bl d m n cyte cells in medium supplemented rhGM-CSF and rhIL-4. T he sec nd step is t exp se t m n - cyte c nditi ned medium mimic r T NF-A. Results There w as n significant difference bserved in dendritic cells yield and purity w hen 5% AB human plasma replace 10% fetal calf serum. Dendritic cells exp sed t m n cyte c nditi ned medium mimic express a higher percentage f restricted mature marker CD83+ and str nger T cell stimulat ry functi n c mparing t exp se t T NF-A. Conclusion We suggest that these mature dendritic cells will be effect ive in viv as adjuvant f r active immun therapy . Key words:Dendrit ic Cell; M n cyte C nditi ned Medium M imic; Mature Regulati n; Immun therapy : 探讨从患者 周血树突状细胞前体培养足够数量的树突状细胞及调控其成熟的方法 完全培养基用5% 人AB 型血浆代替10% 胎牛血清, 培养过程分两阶段, 第一阶段: 从患者 周血 分离出能黏附塑料的单核细胞, 在 GM-CSF+ IL-4 存在条件下培养5~ 7d; 第二阶段, 加入模拟单核细胞 条件性培养液或T NF-A促进树突状细胞分化成熟 5% 人A B 型血浆代替10% 胎牛血清后, 在树 突状细胞的获得率和纯度方面无统计学差异, 模拟单核细胞条件性培养液使树突状细胞表达更高比例