临床级树突状细胞的分离和培养
Large-scale Generation of Mature Clinical Grade Dendritic Cells
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摘要: 目的 探讨从患者外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。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 :从患者外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TNF α促进树突状细胞分化成熟。结果 5 %人AB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的成熟标志CD83+,具有更强的刺激T细胞的能力。结论 通过以上方法培养的成熟树突状细胞将能有效地应用于临床免疫治疗Abstract: Objective To study the method for generation large scale mature dendritic cells from nonproliferation progenitors in patient s blood. Methods The procedure uses 5% AB human plasma in the place of 10% fetal calf serum and involves two steps. The first step is to work on a 5～7 day culture of plastic adherent blood monocyte cells in medium supplemented rhGM CSF and rhIL 4. The second step is to expose to monocyte conditioned medium mimic or TNF α. Results There was no significant difference observed in...
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