Differentiation of natural killer cells from induced pluripotent stem cells under defined, serum- and feeder-free conditions

Kyle B Lupo; Jung-Il Moon; Andrea M Chambers; Sandro Matosevic

doi:10.1016/j.jcyt.2021.05.001

Differentiation of natural killer cells from induced pluripotent stem cells under defined, serum- and feeder-free conditions

Cytotherapy. 2021 Oct;23(10):939-952. doi: 10.1016/j.jcyt.2021.05.001. Epub 2021 Jul 14.

Authors

Kyle B Lupo¹, Jung-Il Moon², Andrea M Chambers¹, Sandro Matosevic³

Affiliations

¹ Department of Industrial and Physical Pharmacy, Purdue University, West Lafayette, Indiana, USA.
² Department of Industrial and Physical Pharmacy, Purdue University, West Lafayette, Indiana, USA; Purdue Institute for Integrative Neuroscience, Purdue University, West Lafayette, Indiana, USA.
³ Department of Industrial and Physical Pharmacy, Purdue University, West Lafayette, Indiana, USA; Purdue Center for Cancer Research, West Lafayette, Indiana, USA. Electronic address: sandro@purdue.edu.

PMID: 34272175
DOI: 10.1016/j.jcyt.2021.05.001

Abstract

Background aims: Traditionally, natural killer (NK) cells are sourced from the peripheral blood of donors-a laborious and highly donor-specific process. Processes for generating NK cells from induced pluripotent stem cells (iPSCs) have demonstrated that it is possible to successfully generate renewable alloreactive NK cells that are not only functional in vivo but can also be genetically engineered for enhanced function. However, poor standardization and cumbersome differentiation procedures suggest that further improvements in the control of the differentiation process are necessary.

Methods: Here the authors evaluated the potential of differentiating NK cells from centrally authenticated iPSCs under entirely chemically defined and serum-free conditions as well as their immunotherapeutic potential, after expansion in feeder-free media, against solid tumors targets. To address limitations of current differentiation approaches, the authors did not utilize feeder or stromal cell layers, TrypLE adaptation or peripheral blood during the differentiation process. The authors also evaluated the feasibility of utilizing centrally authenticated iPSC lines, thus circumventing protocol- and donor-induced variability associated with reprogramming approaches, and characterized these iPSC-NK cells in terms of cytotoxicity, cytokine production and degranulation potential against solid tumor cell lines and patient-derived targets.

Results: Differentiation of iPSCs generated NK cells that were predominantly CD56⁺/CD16⁺/CD3^- and expressed NK activation markers NKG2D, NKp30, NKp44, NKp46 and DNAM-1. These iPSC-NK cells mediated effector functions, including cytotoxicity, degranulation and IFN-γ production, in response to solid tumor targets, including patient-derived cancer cells, and could be cryopreserved and expanded in culture.

Conclusions: The ability to produce NK cells under defined conditions and the functional responses elicited by these iPSC-NK cells suggest that they could represent promising effectors in clinical adoptive transfer settings as a renewable source of donor-independent NK cells for immunotherapy of solid tumors.

Keywords: adoptive transfer; immunotherapy; induced pluripotent stem cells; natural killer cells; solid tumors.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Cell Differentiation
Cell Line, Tumor
Humans
Immunotherapy
Induced Pluripotent Stem Cells*
Killer Cells, Natural

Abstract

Publication types

MeSH terms

Grants and funding