来源于造血干细胞（HSCs）的单核细胞在免疫反应中对癌症起着关键作用。虽然它们是治疗癌症的细胞疗法的有吸引力的来源，但尚未建立起体外扩增的方法。源自多能干细胞（PSC）的分化单核细胞，包括诱导多能干细胞（iPSCs），具有自我更新和多能性，因此可以成为HSC衍生单核细胞的替代来源。为了开发一种用于未来临床应用的iPSC衍生单核细胞的标准化方法，我们旨在控制iPSC群的大小。为此，我们研制了一种含有化学基质的小点盘，用于iPSC支架。放置于该盘中的iPSC仅在点上扩增，并形成相同大小的群。然后，通过向群添加细胞因子将细胞分化为单核细胞。与在覆盖整个表面积的基质盘上培养细胞相比，小点盘极大地减少了单核样细胞生成的变异性。此外，通过调整点的大小和点之间的距离，可获得更多的单核样细胞。iPSC衍生的单核样细胞吞噬癌细胞并分泌促炎细胞因子。细胞还表达Fc受体，并通过相应的抗体对癌细胞进行IgG介导的杀伤。小点盘使得在二维培养中可以控制iPSC群的大小，从而减少功能性单核样细胞的生成变异。使用小点盘产生iPSC衍生的单核样细胞的这种标准化方法也可以促进开发大规模临床应用的自动封闭系统的发展。 版权所有 © 2023 国际细胞与基因疗法学会。Elsevier公司出版。保留所有权利。

Monocytes, derived from hematopoietic stem cells (HSCs), play a pivotal role in the immune response to cancer. Although they are an attractive source of cell therapy for cancer, a method for ex vivo expansion has not yet been established. Monocytes differentiated from pluripotent stem cells (PSCs), including induced pluripotent stem cells (iPSCs), can be an alternative source of HSC-derived monocytes because of their self-renewal and pluripotency. To develop a standardized method for the generation of iPSC-derived monocytes for future clinical applications, we aim to control the size of the iPSC colony.To this end, we developed a plate with multiple dots containing a chemical substrate for the iPSC scaffold. iPSCs placed in the plate expanded only on the dots and created colonies of the same size. The cells were then differentiated into monocytes by adding cytokines to the colonies.The dot plate substantially reduced variability in monocyte-like cell generation when compared with cultivating cells on a plate with the substrate covering the entire surface area. Furthermore, more monocyte-like cells were obtained by adjusting the dot size and the distance between the dots. The iPSC-derived monocyte-like cells phagocytosed cancer cells and secreted proinflammatory cytokines. The cells also expressed Fc receptors and exerted immunoglobulin G-mediated killing of cancer cells with the corresponding antibodies.The dot plate enabled the control of iPSC colony size in two-dimensional culture, which resulted in a reduction in the generation-variation of functional monocyte-like cells. This standardized method for generating iPSC-derived monocyte-like cells using the dot plate could also facilitate the development of an automated closed system on a large scale for clinical applications.Copyright © 2023 International Society for Cell & Gene Therapy. Published by Elsevier Inc. All rights reserved.