Identification, Isolation and in Vitro Expansion of Human and Nonhuman Primate T Stem Cell Memory Cells

Overview

Journal Nat Protoc

Specialties Biology
Pathology
Science

Date 2012 Dec 11

PMID 23222456

Citations 110

Authors

Enrico Lugli

Luca Gattinoni

Alessandra Roberto

Domenico Mavilio

David A Price

Nicholas P Restifo

Mario Roederer

Affiliations

Soon will be listed here.

Abstract

The T cell compartment is phenotypically and functionally heterogeneous; subsets of naive and memory cells have different functional properties, and also differ with respect to homeostatic potential and the ability to persist in vivo. Human stem cell memory T (T(SCM)) cells, which possess superior immune reconstitution and antitumor response capabilities, can be identified by polychromatic flow cytometry on the basis of the simultaneous expression of several naive markers together with the memory marker CD95. We describe here a protocol based on the minimum set of markers required for optimal identification of human and nonhuman primate (NHP) T(SCM) cells with commonly available flow cytometers. By using flow sorters, T(SCM) cells can thereby be isolated efficiently at high yield and purity. With the use of the 5.5-h isolation procedure, depending on the number of cells needed, the sorting procedure can last for 2-15 h. We also indicate multiple strategies for their efficient expansion in vitro at consistent numbers for functional characterization or adoptive transfer experiments.

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