Real-time quantitative PCR analysis of human memory-like NK cells

Human NK cells were purified from healthy whole blood and were differentiated to memory-like NK cells in vitro by culturing for one day in the presence of interleukin-12 (IL-12, 200µg/mL ), interleukin-15 (IL-15, 200µg/mL ) and interleukin-18 (IL-18, 200µg/mL ). After 24hrs media was changed to media containing only inteleukin-15 (IL-15, 200µg/mL). Cell were allowed to remain in culture for 6 days. RNA was extrated using Qiagen RNAEasy Plus Mico Kit. cDNA synthesis and RT-qPCR were perfromed using Qiagen cDNA synthesis kit and Human Glucose Metabolism RT2 Profiler PCR Array. 4 donors were used in gene expression analysis. Donor 4 was selected as the control to compare with samples from donors 1,2, and 3. qPCR gene expression profiling. Memory-like NK cells from 4 donors were used for gene expression analysis. Each donor was run in triplicates. Donors 1,2 and 3 were compared to donor 4.