A specialized tube to make enrichment of specific cell subsets faster and easier (124.3)

Abstract

Abstract Many experimental protocols require the enrichment of specific cell subsets from peripheral blood. RosetteSep™ cell enrichment and standard mononuclear cell (MNC) preparation both involve density gradient centrifugation, which entails slowly layering the sample over the density gradient medium to avoid mixing, and carefully pipetting to remove the enriched cells after centrifugation. Centrifugation must be performed with the brake off to avoid disturbing the enriched cell layer, further lengthening the process. SepMate™, a centrifugation tube with a specialized insert, was developed to allow rapid layering of the sample onto the density gradient medium, and pouring off of the enriched cells after centrifugation, thus simplifying the entire process. When using SepMate™, the cocktail incubation time and centrifugation time could each be shortened to 10 min, making RosetteSep™ cell enrichment even faster. RosetteSep™ enrichments of mononuclear cell subsets using the SepMate™ tubes and protocol gave equivalent purity and recovery of desired cells compared to using the standard RosetteSep™ protocol, and desired cells could be enriched from whole blood in <30 min. Purities of specific cell types were: CD3 T Cells 96 ± 1 (n=5), CD4 T Cells 94 ± 5 (n=3), CD8 T Cells 85 ± 11 (n=4), B Cells 92 ± 6 (n=3), NK Cells 85 ± 5 (n=5), monocytes 68 ± 8 (n = 4). The protocol is easily scalable to process multiple samples simultaneously, and the SepMate™ tube can also be used to prepare MNCs.