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Abstract：
Natural killer (NK) cells hold promise in cancer treatment due to their ability to spontaneously lyse cancer cells. For clinical use, high quantities of pure, functional NK cells are necessary. Combining adherence-based isolation with specialized media showed the unreliability of the isolation method, but demonstrated the superiority of the NK MACS® medium, particularly in suboptimal conditions. Neither human pooled serum, fetal calf serum (FCS), human platelet lysate, nor chemically defined serum replacement could substitute human AB serum. Interleukin (IL-)2, IL-15, IL-21, and combined CD2/NKp46 stimulation were assessed. IL-21 and CD2/NKp46 stimulation increased cytotoxicity, but reduced NK cell proliferation. IL-15 stimulation alone achieved the highest proliferation, but the more affordable IL-2 performed similarly. The RosetteSep™ human NK cell enrichment kit was effective for isolation, but the presence of peripheral blood mononuclear cells (PBMCs) in the culture enhanced NK cell proliferation, despite similar expression levels of CD16, NKp46, NKG2D, and ICAM-1. In line with this, purified NK cells cultured in NK MACS® medium with human AB serum and IL-2 demonstrated high cytotoxicity against primary glioblastoma stem cells.
摘要：
自然杀伤（NK）细胞由于其自发裂解癌细胞的能力而在癌症治疗中具有希望。临床使用，大量的纯净，功能性NK细胞是必需的。将基于粘附的隔离与专门的介质相结合表明隔离方法的不可靠性，但证明了NKMACS®培养基的优越性，特别是在次优条件下。既不是人类汇集的血清，胎牛血清（FCS），人血小板裂解物，化学定义的血清替代也不能替代人AB血清。评估白细胞介素(IL-)2、IL-15、IL-21和组合的CD2/NKp46刺激。IL-21和CD2/NKp46刺激增加细胞毒性，但降低NK细胞增殖。单独的IL-15刺激实现了最高的增殖，但更实惠的IL-2表现类似。RosetteSep™人NK细胞富集试剂盒对分离有效，但是培养物中外周血单核细胞（PBMC）的存在增强了NK细胞的增殖，尽管CD16、NKp46、NKG2D的表达水平相似，和ICAM-1。与此相符,在具有人AB血清和IL-2的NKMACS®培养基中培养的纯化NK细胞表现出对原代成胶质细胞瘤干细胞的高细胞毒性。
