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

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

Author: Yuanyuan XIE ¹ ; Xiaoyan YANG ¹ ; Rong PAN ¹ ; Lixia GAO ² ; Ling YU ³
Author Information

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, Chongqing 400715, 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, Chongqing 402160, China.
3. 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. lingyu12@swu.edu.cn.
Publication Type:Journal Article
Keywords: Co-culture; Human umbilical vein endothelial cell (HUVEC); Migration; Multilayer paper stack; Natural killer cell (NK cell); Tumor spheroid
MeSH: Killer Cells, Natural/immunology*; Coculture Techniques; Spheroids, Cellular; Humans; Paper; Cell Line, Tumor; Microscopy, Confocal
From: Journal of Zhejiang University. Science. B 2024;25(12):1097-1107
CountryChina
Language:English
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 Z-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.