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URL: https://dx.doi.org/10.17504/protocols.io.v87e9zn

Authors: Maria Streltsova, Sofya Erokhina, Leonid Kanevskiy, Dean Lee, William Telford, Elena Kovalenko

Summary: Here we present the simple and effective method for obtaining clones of NK cells, sorted according to a given set of surface markers. The lifespan of clones grown by this method can reach 14 weeks and cloning efficiency is up to 50%, depending on the original NK cell subset. Obtained clones are functionally active; they are able to lyse target cells and produce IFNγ. The principle of method includes single cell sorting and further clonal expansion in the presence of IL-2 and gene-modified K562 feeder cells. Our method allows to expand the progeny of a single NK cell up to 10-20×106 NK cells suitable to study phenotype, proliferative and functional activity of a certain NK cell clone.The work was supported by Russian Science Foundation, grant #16-15-00309.

Affiliations: Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry RAS, Moscow, 117997, Russian Federation, Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry RAS, Moscow, 117997, Russian Federation, Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry RAS, Moscow, 117997, Russian Federation, Center for Childhood Cancer and Blood Disorders, The Research Institute, Nationwide Children’s Hospital, Columbus, OH, USA, National Cancer Institute, National Institute of Health, Bethesda, 20892, USA, Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry RAS, Moscow, 117997, Russian Federation

External URL: https://doi.org/10.1371/journal.pone.0208469

Version: 1

Publication Date: 2018

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