Isolation of functional murine naive and memory T cell subsets in as little as 15 minutes

Abstract Effective cellular immunity requires naïve CD4+ and CD8+ T cells to undergo activation, proliferation and differentiation into various effector cell populations following antigen exposure. CD4+ and CD8+ T cell activation induces the production of cytokines and cytolytic effector proteins which function to coordinate immune responses or directly eliminate infected cells. While most effector T cells become senescent and apoptotic, some effector T cells further differentiate into memory T cells which can provide long lasting immunity. Although the signals that regulate these processes have been widely investigated, continued efforts will eventually construct a definitive model for the role of T cells in adaptive immunity. In many cases, highly purified cells are required for these studies and current methods for their isolation from mice require either lengthy protocols or flow based cell sorting. We have recently developed three new column-free, immunomagnetic cell isolation kits to isolate untouched naïve and memory CD4+ T cells, and naïve CD8+ T cells in as little as 15 minutes. Purities of up to 98% can be achieved as assessed by CD44 and CD62L expression using the manual EasySep™ pour-off method or the fully automated RoboSep™ cell separator. The purified naïve and memory T cells can be immediately used for downstream assays and are fully functional as assessed by in vitro proliferation and cytokine production assays.