Hyaluronan-based hydrogels as versatile tumor-like models: Tunable ECM and stiffness with genipin-crosslinking
Hyaluronan-based hydrogels as versatile tumor-like models: Tunable ECM and stiffness with genipin-crosslinking
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DOI：
10.1002/jbm.a.36899
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发表时间：
2020-02-17
影响因子：
4.9
通讯作者：
Labat, Beatrice
中科院分区：
文献类型：
--
作者：
Bonnesoeur, Sarah;Morin-Grognet, Sandrine;Labat, Beatrice
三维（3D）仿生细胞培养平台提供了细胞在体内自然经历的更真实的微环境。我们开发了一种可调的透明质酸基水凝胶，可以很容易地被修改，以模仿健康或恶性细胞外基质（ECM）。为此，我们用天然存在于肿瘤组织中的粘附多肽（聚-l-赖氨酸，PLL）或ECM蛋白（III型和IV型胶原）预官能化我们的水凝胶，接下来，我们通过与逐渐浓度的京尼平（GNP）交联来调节它们的刚度。然后，我们在用胶质母细胞瘤和乳腺癌细胞测试之前彻底表征了我们的底物，然后用内皮细胞测试。总的来说，我们的水凝胶表现出（a）对于每个预官能化随着GNP浓度增加的刚度和（B）对于PLL处理的有效酶抗性，以及对于IV型胶原蛋白的有效酶抗性，但程度较低。虽然PLL处理的水凝胶不利于任何胶质母细胞瘤细胞系的培养，但它们以刚度依赖性的方式增强了乳腺癌细胞的增殖。与III型胶原相反，IV型胶原预处理的水凝胶支持胶质母细胞瘤细胞的增殖。我们开发的基于HA的3D肿瘤样模型可以通过简单地调整其生化组成和机械特性为各种癌细胞的研究提供有用的平台。
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.