{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:03:24Z","timestamp":1785402204147,"version":"3.56.0"},"reference-count":44,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T00:00:00Z","timestamp":1780444800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100014189","name":"Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, and Forestry","doi-asserted-by":"crossref","award":["322092-4"],"award-info":[{"award-number":["322092-4"]}],"id":[{"id":"10.13039\/501100014189","id-type":"DOI","asserted-by":"crossref"}]},{"award":["322092-4"],"award-info":[{"award-number":["322092-4"]}],"id":[{"id":"https:\/\/ror.org\/00c25dn03","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Ministry of Trade, Industry & Resources","award":["RS-2025-14383159"],"award-info":[{"award-number":["RS-2025-14383159"]}]},{"DOI":"10.13039\/501100000046","name":"National Research Council Canada","doi-asserted-by":"crossref","award":["CGT-807-1"],"award-info":[{"award-number":["CGT-807-1"]}],"id":[{"id":"10.13039\/501100000046","id-type":"DOI","asserted-by":"crossref"}]},{"award":["CGT-807-1"],"award-info":[{"award-number":["CGT-807-1"]}],"id":[{"id":"https:\/\/ror.org\/04mte1k06","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Background\/Objectives: Adoptive transfer of allogeneic natural killer (NK) cells represents a promising off-the-shelf immunotherapy for cancer, offering advantages in safety and availability over autologous T cell therapies. However, generating therapeutically sufficient NK cell numbers remains challenging due to their low frequency in blood sources. Engineered feeder cell co-cultures have enabled substantial expansions of NK cells to clinically relevant doses. Methods: We evaluated the plasma cell leukemia-derived ARH-77 cell line as a feeder for ex vivo NK cell expansion from healthy donor peripheral blood mononuclear cells (PBMCs). Unmodified ARH-77 was compared to K562, followed by engineering both lines to co-express B7-H6 (NKp30 ligand), CD137L (4-1BBL), IL-15, and IL-15R\u03b1 via sequential lentiviral transduction. PBMCs were co-cultured with irradiated feeders in cytokine-supplemented (IL-2, IL-21, and later IL-15) RPMI-1640 or DMEM\/F-12 medium for up to 28 days. Expansion (fold change in CD3\u2212CD56+ cells), purity, surface receptor expression, and cytotoxicity (against K562 targets) were quantified. Results: Unmodified ARH-77 supported significantly greater NK cell expansion than K562 (model-estimated 681-fold vs. 155-fold at week 4 in RPMI; p = 0.0018), with higher purity but comparable cytotoxicity and receptor profiles. Engineered ARH-77 cells achieved robust expansion in RPMI, comparable to that of engineered K562 cells. In optimized DMEM\/F-12 medium, engineered ARH-77 drove superior expansion (up to model-estimated 101,241-fold; 95% CI 46,771\u2013219,146 at week 4), significantly outperforming engineered K562 (4.4-fold greater; 95% CI 1.01 to 18.54; p = 0.0479) while maintaining high purity and equivalent cytotoxicity. Substantial inter-donor variability influenced expansion magnitude, though relative feeder performance remained consistent across donors. Conclusions: Genetically modified ARH-77 feeder cells provide a potent platform for large-scale ex vivo expansion of functional NK cells.<\/jats:p>","DOI":"10.3390\/cancers18111833","type":"journal-article","created":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T09:33:14Z","timestamp":1780479194000},"page":"1833","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of Genetically Modified ARH-77 Feeder Cells for Efficient Expansion of Natural Killer Cells with Potent Anti-Tumor Activity"],"prefix":"10.3390","volume":"18","author":[{"given":"Yu-Jin","family":"Lim","sequence":"first","affiliation":[{"name":"Department of Companion and Laboratory Animal Science, College of Industrial Science, Kongju National University, Yesan 32439, Chungcheongnam-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bryan","family":"Marr","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Microbiology, and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, ON K1H 8M5, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Safa","family":"Ghaziasgar","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Microbiology, and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, ON K1H 8M5, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cheol-Jung","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Companion and Laboratory Animal Science, College of Industrial Science, Kongju National University, Yesan 32439, Chungcheongnam-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yeon-Ju","family":"Baek","sequence":"additional","affiliation":[{"name":"Department of Applied Biotechnology, Kongju National University, Yesan 32439, Chungcheongnam-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Geun-Seop","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Companion and Laboratory Animal Science, College of Industrial Science, Kongju National University, Yesan 32439, Chungcheongnam-do, Republic of Korea"},{"name":"Research Institute for Natural Products, Kongju National University, Yesan 32439, Chungcheongnam-do, Republic of Korea"},{"name":"VaxCell Bio Therapeutics Co., Ltd., Hwasun 58128, Jeollanam-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Je-Jung","family":"Lee","sequence":"additional","affiliation":[{"name":"VaxCell Bio Therapeutics Co., Ltd., Hwasun 58128, Jeollanam-do, Republic of Korea"},{"name":"Department of Hematology-Oncology, Chonnam National University Hwasun Hospital, Hwasun 58128, Jeollanam-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu-Jin","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Applied Biotechnology, Kongju National University, Yesan 32439, Chungcheongnam-do, Republic of Korea"},{"name":"VaxCell Bio Therapeutics Co., Ltd., Hwasun 58128, Jeollanam-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yurim","family":"An","sequence":"additional","affiliation":[{"name":"Department of Applied Biotechnology, Kongju National University, Yesan 32439, Chungcheongnam-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7703-6421","authenticated-orcid":false,"given":"Seung-Hwan","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Microbiology, and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, ON K1H 8M5, Canada"},{"name":"Ottawa Institute of Systems Biology, Centre for Infection, Immunity, and Inflammation, Faculty of Medicine, University of Ottawa, Ottawa, ON K1H 8M5, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0292-8200","authenticated-orcid":false,"given":"Sang-Ki","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Companion and Laboratory Animal Science, College of Industrial Science, Kongju National University, Yesan 32439, Chungcheongnam-do, Republic of Korea"},{"name":"Department of Applied Biotechnology, Kongju National University, Yesan 32439, Chungcheongnam-do, Republic of Korea"},{"name":"Research Institute for Natural Products, Kongju National University, Yesan 32439, Chungcheongnam-do, Republic of Korea"},{"name":"VaxCell Bio Therapeutics Co., Ltd., Hwasun 58128, Jeollanam-do, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1987","DOI":"10.1038\/s41591-025-03640-8","article-title":"Allogeneic NK Cells with a Bispecific Innate Cell Engager in Refractory Relapsed Lymphoma: A Phase 1 Trial","volume":"31","author":"Nieto","year":"2025","journal-title":"Nat. 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