Although primary human hepatocytes (PHHs) are the gold standard in drug efficacy and metabolism studies, long-term survival of PHHs and maintenance of their hepatic function are still challenging. In this study, we focused on the effect of the initial microenvironment on upregulation and long-term preservation of hepatic function of PHHs encapsulated within biodegradable hydrogel systems. PHHs were encapsulated in RGD-functionalized hybrid hydrogels with various degrees of degradability, and their hepatic functionality was analyzed. Regardless of the hydrogel elastic modulus, the combination with nondegradable hydrogels had a predominantly negative effect on the prompt engraftment of PHHs, whereas a degradable hydrogel with intermediate initial degradability was most effective in maintaining hepatic function. Efficient network formation by PHHs and cocultured cells, along with the control of hydrogel degradation, governed the hepatic functionality at an early stage and upon long-term cultivation. Under optimized conditions, expression of genes involved in biological processes such as focal adhesions, cell survival, cytoskeleton formation, and extracellular matrix interactions was significantly higher than that in a control with relatively delayed initial degradation. Thus, we suggest that the orchestrated control of initial cellular remodeling may play an important role in the maintenance of hepatic function in a three-dimensional PHH culture.

中文翻译：

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优化 3D 水凝胶微环境以增强原代人肝细胞的肝功能。

尽管原代人类肝细胞 (PHHs) 是药物疗效和代谢研究的金标准，但 PHHs 的长期存活和维持其肝功能仍然具有挑战性。在这项研究中，我们专注于初始微环境对封装在可生物降解水凝胶系统中的 PHH 的肝功能上调和长期保存的影响。PHHs 被封装在具有不同降解程度的 RGD 功能化混合水凝胶中，并分析了它们的肝脏功能。不管水凝胶弹性模量如何，与不可降解水凝胶的组合对 PHH 的迅速植入具有主要的负面影响，而具有中等初始降解性的可降解水凝胶在维持肝功能方面最有效。PHH 和共培养细胞形成的有效网络，以及水凝胶降解的控制，在早期和长期培养后控制了肝脏功能。在优化条件下，参与黏着斑、细胞存活、细胞骨架形成和细胞外基质相互作用等生物过程的基因表达明显高于初始降解相对延迟的对照。因此，我们建议初始细胞重塑的协调控制可能在三维 PHH 培养中的肝功能维持中发挥重要作用。参与黏着斑、细胞存活、细胞骨架形成和细胞外基质相互作用等生物过程的基因表达明显高于初始降解相对延迟的对照。因此，我们建议初始细胞重塑的协调控制可能在三维 PHH 培养中的肝功能维持中发挥重要作用。参与黏着斑、细胞存活、细胞骨架形成和细胞外基质相互作用等生物过程的基因表达明显高于初始降解相对延迟的对照。因此，我们建议初始细胞重塑的协调控制可能在三维 PHH 培养中的肝功能维持中发挥重要作用。