Comparing the Yield and Cytotoxicity of NK Cells Expanded with Different Feeder Cells.

Overview

Natural killer (NK) cells are ideal candidates for new cancer treatment methods. Roughly 1 million NK cells can be recovered from 10 mL of blood from a healthy donor. Treating one cancer patient would require at least 10 million NK cells per kg body weight collected from many liters of donor blood. Therefore, we need a way to grow NK cells so there are enough for human treatment. We compared NK cell expansion platforms for proliferation yield and cell activation. Blood collected from an arm vein in a healthy volunteer was centrifuged on a ficoll density gradient to isolate peripheral blood mononuclear cells (PBMCs). Magnetic bead depletion of CD14+, CD3+, and CD19+ from PBMCs yielded CD3-/CD56+ NK cells. These were split between feeder cell platforms P1 and P2 (both being cell lines derived from human leukemia patients). After 3 weeks, NK cells co-cultured with P1 and P2 were stained with Trypan Blue and counted using a hemocytometer determining that they yielded 510-fold and 1000-fold proliferation respectively. Using flow cytometry, we determined NK cell surface expression of activating receptors, maturation markers, and trafficking molecules. P1 NK cells exhibited more expression of CD57 (37% vs. 1.5%) indicating high cytotoxic activity and terminal differentiation. P2 cells expressed more of some activating receptors (CD16, NKG2D, and NKp46). Despite these differences, P1 and P2 NK cells displayed similar killing capability against Jurkat T-cell leukemia cells (76.7% and 73.7% killing respectively). Our results suggest that P1 and P2 are both viable NK cell expansion methods.

Competition history

• AJAS 2018 Category not listed

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