Korean J Blood Transfus.  2012 Apr;23(1):28-37.

Ex vivo Expansion of Human Natural Killer Cells from Blood Retained in a Disposable Platelet Apheresis Set

Affiliations
  • 1Research Center for Cancer Immunotherapy, Chonnam National University Hwasun Hospital, Hwasun, Korea. dcho@chonnam.ac.kr
  • 2Department of Laboratory Medicine, Chonnam National University Medical School, Gwangju, Korea.
  • 3Department of Companion & Laboratory Animal Science, Kongju National University, Yesan, Korea.

Abstract

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.

Keyword

Natural killer cell; Ex vivo expansion; Apheresis set
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