Vanhoeijen Robin, Okkelman Irina A, Rogier Nette, Sedlačík Tomáš, Stöbener Daniel D, Devriendt Bert, Dmitriev Ruslan I, Hoogenboom Richard
Supramolecular Chemistry Group, Centre of Macromolecular Chemistry (CMaC), Department of Organic and Macromolecular Chemistry, Faculty of Sciences, Ghent University, Krijgslaan 281-S4, Ghent 9000, Belgium.
Tissue Engineering and Biomaterials Group, Department of Human Structure and Repair, Faculty of Medicine and Health Sciences, Ghent University, C. Heymanslaan 10, Ghent 9000, Belgium.
Biomacromolecules. 2025 Mar 10;26(3):1860-1872. doi: 10.1021/acs.biomac.4c01627. Epub 2025 Feb 3.
The extracellular matrix (ECM) plays a crucial role in organoid cultures by supporting cell proliferation and differentiation. A key feature of the ECM is its mechanical influence on the surrounding cells, directly affecting their behavior. Matrigel, the most commonly used ECM, is limited by its animal-derived origin, batch variability, and uncontrollable mechanical properties, restricting its use in 3D cell-model-based mechanobiological studies. Poly(2-alkyl-2-oxazoline) (PAOx) synthetic hydrogels represent an appealing alternative because of their reproducibility and versatile chemistry, enabling tuning of hydrogel stiffness and functionalization. Here, we studied PAOx hydrogels with differing compressive moduli for their potential to support 3D cell growth. PAOx hydrogels support spheroid and organoid growth over several days without the addition of ECM components. Furthermore, we discovered intestinal organoid epithelial polarity reversion in PAOx hydrogels and demonstrate how the tunable mechanical properties of PAOx can be used to study effects on the morphology and oxygenation of live multicellular spheroids.
细胞外基质（ECM）通过支持细胞增殖和分化，在类器官培养中发挥着关键作用。ECM的一个关键特征是其对周围细胞的机械影响，直接影响细胞行为。基质胶（Matrigel）是最常用的ECM，但因其来源于动物、批次差异以及机械性能不可控等因素而受到限制，这制约了它在基于3D细胞模型的力学生物学研究中的应用。聚（2-烷基-2-恶唑啉）（PAOx）合成水凝胶因其可重复性和多样的化学性质而成为一种有吸引力的替代物，能够调节水凝胶的硬度并实现功能化。在此，我们研究了具有不同压缩模量的PAOx水凝胶支持3D细胞生长的潜力。PAOx水凝胶在不添加ECM成分的情况下，能够在数天内支持球体和类器官的生长。此外，我们发现PAOx水凝胶中肠道类器官上皮极性发生了反转，并展示了如何利用PAOx的可调机械性能来研究其对活的多细胞球体形态和氧合的影响。
