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Review
. 2022 Apr;19(4):460-481.
doi: 10.1038/s41423-021-00808-3. Epub 2022 Jan 5.

Advances in NK cell production

Affiliations
Review

Advances in NK cell production

Fang Fang et al. Cell Mol Immunol. 2022 Apr.

Abstract

Immunotherapy based on natural killer (NK) cells is a promising approach for treating a variety of cancers. Unlike T cells, NK cells recognize target cells via a major histocompatibility complex (MHC)-independent mechanism and, without being sensitized, kill the cells directly. Several strategies for obtaining large quantities of NK cells with high purity and high cytotoxicity have been developed. These strategies include the use of cytokine-antibody fusions, feeder cells or membrane particles to stimulate the proliferation of NK cells and enhance their cytotoxicity. Various materials, including peripheral blood mononuclear cells (PBMCs), umbilical cord blood (UCB), induced pluripotent stem cells (iPSCs) and NK cell lines, have been used as sources to generate NK cells for immunotherapy. Moreover, genetic modification technologies to improve the proliferation of NK cells have also been developed to enhance the functions of NK cells. Here, we summarize the recent advances in expansion strategies with or without genetic manipulation of NK cells derived from various cellular sources. We also discuss the closed, automated and GMP-controlled large-scale expansion systems used for NK cells and possible future NK cell-based immunotherapy products.

Keywords: Genetic manipulation; NK cell lines; NK cells; PBMCs; UCB; iPSCs.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
The sources of NK cells for immunotherapy and strategies of capacity enhancement. Various type of cells, including PBMCs, UCB, iPSCs, and NK cell lines, have been used as the sources to generate NK cells for immunotherapy. Gene modification technologies, differentiation protocols, and novel synthetic reagents have been developed to improve NK cell function
Fig. 2
Fig. 2
At present, the differentiation stage of PSC-NK can be divided into hematopoietic progenitor cell differentiation stage and NK differentiation stage, and the differentiation system can be divided into two systems according to the existence of feeder cells. For feeder system (up panel), some xenogeneic stromal cell lines, such as OP9, M210-B4, or S17, were used to form hematopoietic progenitor cells (HPCs). However, HPCs can also be obtain through spin embryoid bodies (EBs) with the help of the combination of cytokines (BMP4, SCF, and VEGF) and small molecules (Y-27632). Subsequently, HPCs were co-incubated with MS-5, AFT024 or EL08-1D2, accompanied by the presence of cytokines (IL-7, IL-15, SCF, Flt3L, and IL-3 only first week), finally to generate PSC-NK. For feeder-free system (bottom panel), two differentiation stage did not involve xenogeneic stromal cell lines, and the induction was completed under the combination of cytokines
Fig. 3
Fig. 3
“Unattended” intelligent NK cell factory with “off-the-shelf/customized/self-supporting” capabilities. It is expected that, the NK cells manufacturing just likes an assemble-line, composed of three steps: (1) Starting cell processing and storage, includes starting cell selection, separation and cryopreservation; (2) Expansion and function improvement, including NK cell culture, gene engineering, quality control system and packaging; (3) Storage, the products are cryopreserved. During the manufacturing process, an quality control system involves automatic sampling, test result evaluation and intelligence adjustment of the production process is interacted with the monitor and control system. All the information of the samples, tests and processes running parameters are monitored and controlled by the Monitor and control system
Fig. 4
Fig. 4
Donor and recipient matching system. The NK cell biomarkers from each donor and the characteristic from each recipient were assessed. The matching search engine selects the best matching donor NK cells for an recipient by comparing the NK cell biomarkers from donors and the tumor biomarkers from recipient, and provides an NK cell manufacture process by selecting the available enhancing strategies such as gene modification and arming, to achieve a best predicted efficacy

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