Dong Yixiao, Qian Shuyi, Wang Xuechun, Zhang Wang, Lu Weisheng, Qu Ju, Cui Meihua, Chen Linzhi, Zhao Yingshuai, Gao Yuehua, Giomo Monica, Urciuolo Anna, Feng Jian, Zheng Yijun, Jiang Biao, Shen Ruling, Zhu Xianmin, Elvassore Nicola
Shanghai Clinical Research and Trial Center, Shanghai Institute for Advanced Immunochemical Studies (SIAIS), ShanghaiTech University, Shanghai, 201210, China.
Shanghai Academy of Sciences & Technology Institute of Model Animals Transformation, Shanghai, 201203, China.
Mater Today Bio. 2025 Feb 19;31:101602. doi: 10.1016/j.mtbio.2025.101602. eCollection 2025 Apr.
The mechanical properties and physical confinement of the extracellular matrix (ECM) are crucial roles in regulating tumor growth and progression. Extensive efforts have been dedicated to replicating the physical characteristics of tumor tissue by developing two-dimensional (2D) and three-dimensional (3D) models. However, it remains a significant challenge to modulate the local microenvironment around the specific cells according to the culture progress. In this study, we develop a 3D culture platform for multicellular lung cancer spheroids using a gelatin-based hydrogel with adjustable density and stiffness. Then, by utilizing a two-photon mediated bioprinting technique, we construct 3D confining microstructures with micrometer accuracy to enclose the selected spheroids within the hydrogel matrix. Diverse transcriptional profilings of cells are observed in response to the increased ECM density and stiffness compared to the additional confining stress. In addition, changed confining stress can regulate the tumor cells with contrary impacts on the cell cycle-related pathways. Our model not only allows for modifications to the mechanical microenvironment of the overall matrix but also facilitates localized adjustments throughout the culture evolution. This approach serves as a valuable tool for investigating tumor progression and understanding cell-ECM interactions.
细胞外基质（ECM）的力学性能和物理限制在调节肿瘤生长和进展中起着关键作用。人们通过开发二维（2D）和三维（3D）模型，付出了大量努力来复制肿瘤组织的物理特征。然而，根据培养进程调节特定细胞周围的局部微环境仍然是一项重大挑战。在本研究中，我们使用具有可调密度和刚度的明胶基水凝胶开发了一种用于多细胞肺癌球体的3D培养平台。然后，通过利用双光子介导的生物打印技术，我们构建了具有微米精度的3D限制微结构，将选定的球体包裹在水凝胶基质中。与额外的限制应力相比，观察到细胞对增加的ECM密度和刚度有不同的转录谱。此外，改变的限制应力可以调节肿瘤细胞，对细胞周期相关途径产生相反的影响。我们的模型不仅允许对整个基质的力学微环境进行修改，还便于在整个培养过程中进行局部调整。这种方法是研究肿瘤进展和理解细胞与ECM相互作用的宝贵工具。
