Chen Yu-Hsu, Ku Yuan-Hao, Wang Kuo-Cheng, Chiang Hung-Chi, Hsu Yu-Pao, Cheng Ming-Te, Wang Ching-Shuen, Wee Yinshen
Department of Orthopedics, Taoyuan General Hospital, Ministry of Health and Welfare, Taoyuan City 330, Taiwan.
Department of General & MIS Surgery, Taoyuan General Hospital, Ministry of Health and Welfare, Taoyuan City 330, Taiwan.
Gels. 2022 Feb 27;8(3):149. doi: 10.3390/gels8030149.
The generation of hepatic spheroids is beneficial for a variety of potential applications, including drug development, disease modeling, transplantation, and regenerative medicine. Natural hydrogels are obtained from tissues and have been widely used to promote the growth, differentiation, and retention of specific functionalities of hepatocytes. However, relying on natural hydrogels for the generation of hepatic spheroids, which have batch to batch variations, may in turn limit the previously mentioned potential applications. For this reason, we researched a way to establish a three-dimensional (3D) culture system that more closely mimics the interaction between hepatocytes and their surrounding microenvironments, thereby potentially offering a more promising and suitable system for drug development, disease modeling, transplantation, and regenerative medicine. Here, we developed self-assembling and bioactive hybrid hydrogels to support the generation and growth of hepatic spheroids. Our hybrid hydrogels (PC4/Cultrex) inspired by the sandcastle worm, an Arg-Gly-Asp (RGD) cell adhesion sequence, and bioactive molecules derived from Cultrex BME (Basement Membrane Extract). By performing optimizations to the design, the PC4/Cultrex hybrid hydrogels can enhance HepG2 cells to form spheroids and express their molecular signatures (e.g., , , , , , , and ). Our study demonstrated that this hybrid hydrogel system offers potential advantages for hepatocytes in proliferating, differentiating, and self-organizing to form hepatic spheroids in a more controllable and reproducible manner. In addition, it is a versatile and cost-effective method for 3D tissue cultures in mass quantities. Importantly, we demonstrate that it is feasible to adapt a bioinspired approach to design biomaterials for 3D culture systems, which accelerates the design of novel peptide structures and broadens our research choices on peptide-based hydrogels.
肝球体的生成有利于多种潜在应用，包括药物开发、疾病建模、移植和再生医学。天然水凝胶是从组织中获得的，已被广泛用于促进肝细胞的生长、分化和特定功能的保留。然而，依赖具有批次间差异的天然水凝胶来生成肝球体，可能会反过来限制上述潜在应用。因此，我们研究了一种建立三维（3D）培养系统的方法，该系统能更紧密地模拟肝细胞与其周围微环境之间的相互作用，从而有可能为药物开发、疾病建模、移植和再生医学提供一个更有前景且合适的系统。在这里，我们开发了自组装和生物活性混合水凝胶来支持肝球体的生成和生长。我们的混合水凝胶（PC4/Cultrex）受到沙堡蠕虫的启发，具有精氨酸 - 甘氨酸 - 天冬氨酸（RGD）细胞粘附序列以及源自Cultrex BME（基底膜提取物）的生物活性分子。通过对设计进行优化，PC4/Cultrex混合水凝胶可以增强HepG2细胞形成球体并表达其分子特征（例如， ， ， ， ， ，和 ）。我们的研究表明，这种混合水凝胶系统在使肝细胞以更可控和可重复的方式增殖、分化和自组织形成肝球体方面具有潜在优势。此外，它是一种用于大量3D组织培养的通用且经济高效的方法。重要的是，我们证明了采用仿生方法设计用于3D培养系统的生物材料是可行的，这加速了新型肽结构的设计，并拓宽了我们对基于肽的水凝胶的研究选择。
