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Updated: Aug 1, 2026
Natural Killer NK and CAR-NK Cell Expansion Method using Membrane Bound-IL-21-Modified B Cell Line
Published on: February 8, 2022
Generation and Functional Characterization of Anti-CD19 Chimeric Antigen Receptor-Natural Killer Cells from Human
Phatchanat Klaihmon1, Xing Kang1, Surapol Issaragrisil1,2,3
1Siriraj Center of Excellence for Stem Cell Research, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
Abstract:
Natural killer (NK) cells are a part of innate immunity that can be activated rapidly in response to malignant transformed cells without prior sensitization. Engineering NK cells to express chimeric antigen receptors (CARs) allows them to be directed against corresponding target tumor antigens. CAR-NK cells are regarded as a promising candidate for cellular immunotherapy alternatives to conventional CAR-T cells, due to the relatively low risk of graft-versus-host disease and safer clinical profile. Human induced pluripotent stem cells (iPSCs) are a promising renewable cell source of clinical NK cells. In the present study, we successfully introduced a third-generation CAR targeting CD19, which was validated to have effective signaling domains suitable for NK cells, into umbilical cord blood NK-derived iPSCs, followed by a single-cell clone selection and thorough iPSC characterization. The established single-cell clone of CAR19-NK/iPSCs, which is highly desirable for clinical application, can be differentiated using serum- and feeder-free protocols into functional CAR19-iNK-like cells with improved anti-tumor activity against CD19-positive hematologic cancer cells when compared with wild-type (WT)-iNK-like cells. With the feasibility of being an alternative source for off-the-shelf CAR-NK cells, a library of single-cell clones of CAR-engineered NK/iPSCs targeting different tumor antigens may be created for future clinical application.
Insights
Engineered natural killer (NK) cells offer a safer cellular immunotherapy. Researchers created chimeric antigen receptor (CAR)-NK cells from stem cells, showing enhanced anti-tumor activity against blood cancers.
Area of Science:
- Immunology
- Cellular Therapy
- Biotechnology
Background:
- Natural killer (NK) cells are crucial for innate immunity, rapidly targeting malignant cells without prior sensitization.
- Chimeric antigen receptor (CAR)-engineered NK cells represent a promising cellular immunotherapy alternative to CAR-T cells, offering a potentially safer clinical profile and lower risk of graft-versus-host disease.
- Human induced pluripotent stem cells (iPSCs) provide a renewable and scalable source for generating clinical-grade NK cells.
Purpose of the Study:
- To engineer human induced pluripotent stem cells (iPSCs) to produce chimeric antigen receptor (CAR)-NK cells targeting CD19.
- To characterize the established CAR19-NK/iPSC single-cell clone and its differentiated iNK-like cells.
- To evaluate the anti-tumor efficacy of CAR19-iNK-like cells against CD19-positive hematologic cancer cells.
Main Methods:
- Introduction of a third-generation CAR targeting CD19 into umbilical cord blood NK-derived iPSCs.
- Single-cell clone selection and comprehensive characterization of the CAR-engineered iPSCs (CAR19-NK/iPSCs).
- Differentiation of CAR19-NK/iPSCs into CAR19-iNK-like cells using serum- and feeder-free protocols and assessment of their anti-tumor activity.
Main Results:
- Successful generation and characterization of a single-cell clone of CAR19-NK/iPSCs.
- Differentiation into functional CAR19-iNK-like cells with enhanced anti-tumor activity against CD19-positive hematologic cancer cells compared to wild-type iNK-like cells.
- Demonstrated feasibility of using iPSCs as a source for off-the-shelf CAR-NK cells.
Conclusions:
- CAR19-NK/iPSCs represent a valuable cellular product for clinical applications in hematologic malignancies.
- iPSC-derived CAR-NK cells offer a scalable and potentially off-the-shelf alternative for cancer immunotherapy.
- This platform allows for the creation of a library of CAR-engineered NK/iPSCs targeting diverse tumor antigens for future therapeutic development.
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