Co-culture of natural killer cells and tumor spheroids on a heterogeneous multilayer paper stack

Yuanyuan XIE; Xiaoyan YANG; Rong PAN; Lixia GAO; Ling YU

doi:10.1631/jzus.B2300617

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Research Article | Updated：2024-12-27

Co-culture of natural killer cells and tumor spheroids on a heterogeneous multilayer paper stack
Co-culture of natural killer cells and tumor spheroids on a heterogeneous multilayer paper stack
自然杀伤细胞与肿瘤球在多层纸基堆叠支架的共培养
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology) Vol. 25, Issue 12, Pages: 1097-1107(2024)
Affiliations：

1.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, Chongqing400715, China
2.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, Chongqing402160, China
Funds：
DOI：10.1631/jzus.B2300617
CLC：
CSTR：
Published： 15 December 2024 ，

Received： 03 September 2023 ，

Revised： 04 March 2024 ，
Accepted：

谢媛媛,杨晓艳,潘茸等.自然杀伤细胞与肿瘤球在多层纸基堆叠支架的共培养[J].浙江大学学报（英文版）（B辑：生物医学和生物技术）,2024,25(12):1097-1107.

YUANYUAN XIE, XIAOYAN YANG, RONG PAN, et al. Co-culture of natural killer cells and tumor spheroids on a heterogeneous multilayer paper stack. [J]. Journal of zhejiang university-science b (biomedicine & biotechnology), 2024, 25(12): 1097-1107.
谢媛媛,杨晓艳,潘茸等.自然杀伤细胞与肿瘤球在多层纸基堆叠支架的共培养[J].浙江大学学报（英文版）（B辑：生物医学和生物技术）,2024,25(12):1097-1107. DOI： 10.1631/jzus.B2300617.

YUANYUAN XIE, XIAOYAN YANG, RONG PAN, et al. Co-culture of natural killer cells and tumor spheroids on a heterogeneous multilayer paper stack. [J]. Journal of zhejiang university-science b (biomedicine & biotechnology), 2024, 25(12): 1097-1107. DOI： 10.1631/jzus.B2300617.

摘要

多层纸基堆叠支架细胞培养体系作为一种三维（3D）体外细胞培养平台，已被用于评估抗肿瘤药物的疗效。本研究提出了一种基于多层纸基堆叠支架的共培养模型，用于模拟自然杀伤细胞（NK细胞）从内皮层向肿瘤细胞球体迁移并发起攻击的过程。该共培养体系由三层结构组成：底层为肿瘤细胞球体层；中间层为模拟肿瘤间质的侵袭层；顶层为内皮细胞层。NK-92细胞悬浮于培养体系上方的培养基中。经过2天的共培养后，我们观察到NK细胞与肿瘤细胞球体之间发生相互作用。此外，共聚焦显微镜检测结果发现NK-92细胞能够穿过内皮层，在肿瘤细胞球体内部不同深度处浸润，形成肿瘤浸润NK细胞（TINKs），这表明该共培养模型成功模拟了NK细胞向实体肿瘤组织浸润的过程。该多层纸基共培养体系在涉及肿瘤细胞与免疫细胞相互作用的模型研究中具有应用潜力，为探索两种细胞间动力学相互作用提供了条件。

Abstract

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.

关键词

多层堆叠纸基共培养肿瘤球人脐静脉内皮细胞人自然杀伤细胞迁移

Keywords

Multilayer paper stackCo-cultureTumor spheroidHuman umbilical vein endothelial cell (HUVEC)Natural killer cell (NK cell)Migration

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