K562细胞
白细胞介素12
白细胞介素21
Janus激酶3
淋巴因子激活杀伤细胞
外周血单个核细胞
细胞生物学
NK-92
白细胞介素15
生物
细胞培养
分子生物学
细胞毒性T细胞
免疫学
化学
T细胞
体外
细胞因子
免疫系统
白细胞介素
生物化学
遗传学
作者
Minh‐Trang Thi Phan,Seung Hwan Lee,Sang‐Ki Kim,Duck Cho
出处
期刊:Methods in molecular biology
日期:2016-01-01
卷期号:: 167-174
被引量:32
标识
DOI:10.1007/978-1-4939-3684-7_14
摘要
Natural killer (NK) cells can be expanded upon activation by proliferative cytokines (such as IL-2 and IL-15). The NK cell expansion can be greatly enhanced by proteins from feeder cells such as tumor cell lines or PBMCs. Therefore, coculture systems of irradiated feeder cells and NK cells in media containing IL-2 and IL-15 have been developed to generate large numbers of NK cells, although NK cell expansion protocol using anti-CD3 antibody (OKT-3) without feeder cells has also been developed. Commonly used feeder cell lines are RPMI8866, Epstein-Barr lymphoblastoid cell line (EBV-LCL), and K562. Stimulation with NK-sensitive K562 cells is known to augment NK cell proliferation to IL-2, IL-15, and IL-21 in combination.Recently, remarkable NK cell-expansion rates are achieved when genetically engineered (GE) feeder cells are used. Dr. Dario Campana's group found that membrane-bound IL-15 and 4-1BBL, coexpressed by K562 cells, acted synergistically to augment K562-specific NK stimulatory capacity, resulting in vigorous expansion of peripheral blood CD56(+) CD3(-) NK cells without concomitant growth of T lymphocytes. Here, we describe an in vitro expansion method of human NK cells among PBMCs by coculturing with GE_K562 cells.
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