Rapid Transduction and Expansion of Transduced T Cells with Maintenance of Central Memory Populations

Mary S. Pampusch, Kumudhini Preethi Haran, Geoffrey T. Hart, Eva G. Rakasz, Aaron K. Rendahl, Edward A. Berger, Elizabeth Connick, Pamela J. Skinner

Research output: Contribution to journalArticlepeer-review

22 Scopus citations


Chimeric antigen receptor (CAR)-T cells show great promise in treating cancers and viral infections. However, most protocols developed to expand T cells require relatively long periods of time in culture, potentially leading to progression toward populations of terminally differentiated effector memory cells. Here, we describe in detail a 9-day protocol for CAR gene transduction and expansion of primary rhesus macaque peripheral blood mononuclear cells (PBMCs). Cells produced and expanded with this method show high levels of viability, high levels of co-expression of two transduced genes, retention of the central memory phenotype, and sufficient quantity for immunotherapeutic infusion of 1–2 × 108 cells/kg in a 10 kg rhesus macaque. This 9-day protocol may be broadly used for CAR-T cell and other T cell immunotherapy approaches to decrease culture time and increase maintenance of central memory populations.

Original languageEnglish (US)
Pages (from-to)1-10
Number of pages10
JournalMolecular Therapy - Methods and Clinical Development
StatePublished - Mar 13 2020

Bibliographical note

Publisher Copyright:
© 2019 The Author(s)


  • CAR-T cells
  • PBMC
  • central memory
  • expansion
  • retrovirus
  • rhesus macaque
  • transduction


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