Kunkanjanawan Hataiwan, Somredngan Sirilak, Kunkanjanawan Tanut, Wongtrakoongate Patompon, Wongsakmanee Wannida, Khemarangsan Veerapol, Masuyama Jun-Ichi, Parnpai Rangsun
Embryo Technology and Stem Cell Research Center, School of Biotechnology, Suranaree University of Technology, Nakhon Ratchasima, Thailand; Medeze Research and Development Co., Ltd., Nakhon Pathom, Thailand.
Medeze Group Public Company Limited, Nakhon Pathom, Thailand.
Cytotherapy. 2025 Feb 24. doi: 10.1016/j.jcyt.2025.02.006.
Growing research in the immunotherapy field has shed light on the adoptive cell transfer efficacy of ex vivo expanded natural killer (NK) cells in cancer treatment. In accordance with the advantages of using cord blood as a source of hematopoietic cells, this study aimed to establish a sustainable supply of NK cells by developing a simple method for expanding cord blood-derived NK (CBNK) cells without using feeders or cell-sorting processes. To achieve this aim, culture strategies that stimulate the proliferation of NK cells from hematopoietic stem and progenitor cells (HSPCs) would result in a high purity of CBNK cells. Here, we first compared the potential of lipopolysaccharide (LPS) and the Notch signaling agonist Yhhu-3792 to promote CBNK cell proliferation in cytokine-based conditions before applying these findings to a large-scale expansion platform. Overall, we reveal that the presence of LPS at 1 µg/mL during the first week of a 21-day expansion protocol resulted in an average total nucleated cell (TNC) count of 1.68 ± 2.92 × 10, with 92.09 ± 3.47% of the expanded cells being NK cells (n = 5). All subsequent analyses demonstrated that the expanded CBNK cells produced by this procedure are phenotypically and functionally competent NK cells. Collectively, this study developed a simple cytokine-based and cell-sorting-free method for the large-scale expansion of CBNK cells.
免疫治疗领域日益增多的研究揭示了体外扩增的自然杀伤（NK）细胞在癌症治疗中的过继性细胞转移疗效。鉴于使用脐带血作为造血细胞来源的优势，本研究旨在开发一种简单的方法来扩增脐带血来源的NK（CBNK）细胞，无需使用饲养细胞或细胞分选过程，从而建立CBNK细胞的可持续供应。为实现这一目标，刺激造血干细胞和祖细胞（HSPCs）中NK细胞增殖的培养策略将产生高纯度的CBNK细胞。在此，我们首先比较了脂多糖（LPS）和Notch信号激动剂Yhhu-3792在基于细胞因子的条件下促进CBNK细胞增殖的潜力，然后将这些发现应用于大规模扩增平台。总体而言，我们发现，在21天扩增方案的第一周添加1µg/mL的LPS，平均总核细胞（TNC）计数为1.68±2.92×10，扩增细胞中92.09±3.47%为NK细胞（n = 5）。所有后续分析表明，通过该程序扩增的CBNK细胞在表型和功能上均为有功能的NK细胞。总之，本研究开发了一种简单的基于细胞因子且无需细胞分选的方法，用于大规模扩增CBNK细胞。
