Xie Yuanyuan, Yang Xiaoyan, Pan Rong, Gao Lixia, Yu Ling
Key Laboratory of Luminescence Analysis and Molecular Sensing, Ministry of Education, Institute for Clean Energy and Advanced Materials, School of Materials and Energy, Southwest University, Chongqing 400715, China.
National & Local Joint Engineering Research Center of Targeted and Innovative Therapeutics, College of Pharmacy & International Academy of Targeted Therapeutics and Innovation, Chongqing University of Arts and Sciences, Chongqing 402160, China.
J Zhejiang Univ Sci B. 2024 Dec 15;25(12):1097-1107. doi: 10.1631/jzus.B2300617.
Multilayer paper-based cell culture, as an in vitro three-dimensional (3D) cell culture method, has been frequently used to research drug bioavailability, therapeutic efficacy, and dose-limiting toxicity in malignant tumors. This paper proposes a heterogenous multilayer paper stacking co-culture system to establish a model of natural killer (NK) cells moving through the endothelium layer and attacking tumor spheroids. This system consists of three layers: a bottom tumor-spheroid layer, a middle invasion layer, and a top endothelium layer. NK-92 cells were placed in the supernatant on top of the three layers. After two days of co-culture, the attack of tumor spheroids by NK cells was observed. We additionally examined the infiltration of NK-92 cells within the tumor spheroids at different -axis depths using a confocal microscope, and the results suggested that the system successfully realizes NK cells traveling cross the endothelium layer to form tumor-infiltrating NK cells (TINKs). The potential application of multilayer paper for co-culture models involving cancer cells and immune cells holds great promise for exploring the interaction dynamics of these two cell types.
多层纸质细胞培养作为一种体外三维（3D）细胞培养方法，已被频繁用于研究恶性肿瘤中的药物生物利用度、治疗效果和剂量限制性毒性。本文提出了一种异质多层纸堆叠共培养系统，以建立自然杀伤（NK）细胞穿过内皮细胞层并攻击肿瘤球体的模型。该系统由三层组成：底部肿瘤球体层、中间侵袭层和顶部内皮细胞层。将NK-92细胞置于三层顶部的上清液中。共培养两天后，观察到NK细胞对肿瘤球体的攻击。我们还使用共聚焦显微镜检查了NK-92细胞在不同轴深度的肿瘤球体内的浸润情况，结果表明该系统成功实现了NK细胞穿过内皮细胞层形成肿瘤浸润性NK细胞（TINKs）。多层纸在涉及癌细胞和免疫细胞的共培养模型中的潜在应用，对于探索这两种细胞类型的相互作用动态具有很大的前景。
