Normandie Université, INSERM, U1234, Faculté de Médecine et Pharmacie, UNIROUEN, Rouen, France.
Normandie Université, PBS UMR 6270, UFR de Sciences et Techniques, FR3038, UNIROUEN, INSA Rouen, CNRS, Evreux Cedex, France.
J Biomed Mater Res A. 2020 May;108(5):1256-1268. doi: 10.1002/jbm.a.36899. Epub 2020 Feb 17.
Three-dimensional (3D) biomimetic cell culture platforms offer more realistic microenvironments that cells naturally experience in vivo. We developed a tunable hyaluronan-based hydrogels that could easily be modified to mimic healthy or malignant extracellular matrices (ECMs). For that, we pre-functionalized our hydrogels with an adhesive polypeptide (poly-l-lysine, PLL) or ECM proteins (type III and type IV collagens), naturally present in tumorous tissues, and next, we tuned their stiffness by crosslinking with gradual concentrations of genipin (GnP). Then, we thoroughly characterized our substrates before testing them with glioblastoma and breast cancer cells, and thereafter with endothelial cells. Overall, our hydrogels exhibited (a) increasing stiffness with GnP concentration for every pre-functionalization and (b) efficient enzyme resistance with PLL treatment, and also with type IV collagen but to a lesser extent. While PLL-treated hydrogels were not favorable to the culture of any glioblastoma cell lines, they enhanced the proliferation of breast cancer cells in a stiffness-dependent manner. Contrary to type III collagen, type IV collagen pre-treated hydrogels supported the proliferation of glioblastoma cells. The as-desired HA-based 3D tumor-like models we developed may provide a useful platform for the study of various cancer cells by simply tuning their biochemical composition and their mechanical properties.
三维（3D）仿生细胞培养平台提供了更接近细胞在体内自然经历的真实微环境。我们开发了一种可调节的透明质酸基水凝胶，可轻松修饰以模拟健康或恶性细胞外基质（ECM）。为此，我们用一种粘附多肽（聚-L-赖氨酸，PLL）或天然存在于肿瘤组织中的 ECM 蛋白（III 型和 IV 型胶原）对水凝胶进行预功能化，然后用不同浓度的京尼平（GnP）交联来调节其硬度。之后，我们对基质进行了全面的特性分析，然后用神经胶质瘤和乳腺癌细胞进行了测试，之后又用内皮细胞进行了测试。总体而言，我们的水凝胶表现出：（a）对于每种预功能化，随着 GnP 浓度的增加，硬度逐渐增加；（b）PLL 处理以及 IV 型胶原处理具有有效的酶抗性，但程度较小。虽然 PLL 处理的水凝胶不利于任何神经胶质瘤细胞系的培养，但它们以硬度依赖的方式促进了乳腺癌细胞的增殖。与 III 型胶原相反，预处理的 IV 型胶原水凝胶支持神经胶质瘤细胞的增殖。我们开发的基于所需透明质酸的 3D 类肿瘤模型可通过简单调节其生化组成和机械性能，为研究各种癌细胞提供有用的平台。
