3D in vitro co-culture disc for assessing cancer invasiveness in the tumor stroma microenvironment.

IF 3

In vitro models Pub Date : 2026-02-17 eCollection Date: 2026-02-01 DOI:10.1007/s44164-026-00106-0

Weiyou Sun, Yutaka Kikuchi, Haruko Takahashi

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Abstract

Purpose: Cancer cell invasion into the stroma is a key step in tumor progression, yet many in vitro models fail to recapitulate stromal architecture or permit quantitative analysis along the thickness (Z) dimension. This study aimed to develop a simple 3D co-culture disc system that maintains fibroblast-rich gels without shrinkage and enables quantitative evaluation of epithelial cancer cell invasion.

Methods: A thin plastic disc with a central opening was used to support collagen gels containing normal human lung fibroblasts (NHLFs). Noncancerous MCF-10 A and cancer cell lines with different invasive capacities (MCF-7, MDA-MB-231) were seeded on the gel surface. Gel contraction, fibroblast morphology, and invasion depth were assessed using fluorescence labeling and confocal laser scanning microscopy (CLSM). Transforming growth factor-β1 (TGF-β1) was applied to test responsiveness to environmental stimuli.

Results: Fibroblast-rich gels contracted uniaxially along the Z-axis but remained laterally stable within the disc, enabling long-term co-culture. Cancer cells showed distinct invasion profiles, with MDA-MB-231 invading significantly deeper than MCF-10 A or MCF-7. Invading cells frequently localized near αSMA-high myofibroblast-like fibroblasts. TGF-β treatment increased fibroblast contractility and further enhanced cancer cell invasion, demonstrating that the system quantitatively detects stromal activation and invasion-promoting cues.

Conclusion: The disc-based 3D co-culture system offers a simple and physiologically relevant platform for quantifying cancer cell invasion and fibroblast activation. By supporting fibroblast-rich stromal environments, resolving Z-axis invasion, and responding to external stimuli such as TGF-β, this system provides a practical tool for studying tumor-stroma interactions and may be applicable to patient-derived cells in future studies.

Supplementary information: The online version contains supplementary material available at 10.1007/s44164-026-00106-0.

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三维体外共培养盘评估肿瘤间质微环境侵袭性。

目的：癌细胞侵入间质是肿瘤进展的关键步骤，然而许多体外模型无法重现间质结构或允许沿厚度(Z)维度进行定量分析。本研究旨在开发一种简单的3D共培养光盘系统，该系统可以维持富含成纤维细胞的凝胶而不收缩，并能够定量评估上皮癌细胞的侵袭。方法：采用带中心开口的薄塑料盘支撑含有正常人肺成纤维细胞（NHLFs）的胶原凝胶。将非癌性mcf - 10a和具有不同侵袭能力的癌细胞（MCF-7、MDA-MB-231）播种在凝胶表面。使用荧光标记和共聚焦激光扫描显微镜（CLSM）评估凝胶收缩、成纤维细胞形态和浸润深度。应用转化生长因子-β1 （TGF-β1）检测对环境刺激的反应性。结果：富成纤维细胞凝胶沿z轴单向收缩，但在椎间盘内保持横向稳定，可以长期共培养。癌细胞表现出不同的侵袭特征，MDA-MB-231的侵袭深度明显高于mcf - 10a或MCF-7。入侵细胞通常定位于α - sma含量高的肌成纤维细胞样成纤维细胞附近。TGF-β处理增加了成纤维细胞的收缩能力，进一步增强了癌细胞的侵袭，表明该系统定量检测基质激活和侵袭促进线索。结论：基于光盘的三维共培养系统为定量癌细胞侵袭和成纤维细胞活化提供了一种简单、生理相关的平台。通过支持富含成纤维细胞的基质环境，解决z轴侵袭，响应TGF-β等外部刺激，该系统为研究肿瘤-基质相互作用提供了一个实用的工具，并可能在未来的研究中适用于患者来源的细胞。补充信息：在线版本包含补充资料，提供地址为10.1007/s44164-026-00106-0。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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In vitro models

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