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KMID : 0387720120230010028
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Korean Journal of Blood Transfusion
2012 Volume.23 No. 1 p.28 ~ p.37
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Ex vivo Expansion of Human Natural Killer Cells from Blood Retained in a Disposable Platelet Apheresis Set
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Shin Dong-Jun
Cho Duck
Jang Youn-Young
Park Ji-Yun
Kim Ju-Sun
Kim Sang-Ki
Lee Je-Jung
Jang Min-Joong
Shin Myung-Geun
Ryang Dong-Wook
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Abstract
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Background: Natural killer cells expanded from human peripheral blood (PB) have been used in cancer immunotherapy research. Although most research teams have access to human PB, it is necessary to find a source of blood that can be easily obtained. We have tested the possibility of using blood retained in a disposable platelet apheresis set as an alternative source, with special interest in expansion of NK cells for use in cancer immunotherapy research.
Methods: For expansion of NK cells, peripheral blood mononuclear cells (PBMCs) were isolated from an MCS+ platelet apheresis kit (Haemonetics, Braintree, USA) and PB from the same donor (n=7) and co-cultured with 100-Gy gamma ray-irradiated K562 cells expressing the 4-1BB ligand and membrane-bound IL-15 for three weeks in RPMI1640 medium in the presence of IL-2 and IL-15. Cytotoxicity was measured using WST-1 at 1:1, 2:1, and 4:1 effector-to-target (E:T) ratios for a period of four hours.
Results: Mean rate of expansion of NK cells was 1,097-fold and their purity was 94.4% from blood retained in a disposable platelet apheresis set; mean rate of expansion of NK cells was 953-fold and their purity was 92.0% from PB after a period of three weeks. No differences in cytotoxicity against K562, 697, Raji, and RPMI8226 were observed between NK cells expanded from two blood sources.
Conclusion: Blood retained in a disposable platelet apheresis set is a useful and convenient source for expansion of NK cells for use in cancer immunotherapy research.
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KEYWORD
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Natural killer cell, Ex vivo expansion, Apheresis set
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