Department of Biomedical Engineering, University of Illinois at Chicago, Chicago, Illinois.
Department of Biomedical Engineering, University of Illinois at Chicago, Chicago, Illinois.
Cell Mol Gastroenterol Hepatol. 2023;15(5):1147-1160. doi: 10.1016/j.jcmgh.2023.01.013. Epub 2023 Feb 2.
Several liver diseases (eg, hepatitis B/C viruses, alcoholic/nonalcoholic fatty liver, malaria, monogenic diseases, and drug-induced liver injury) significantly impact global mortality and morbidity. Species-specific differences in liver functions limit the use of animals to fully elucidate/predict human outcomes; therefore, in vitro human liver models are used for basic and translational research to complement animal studies. However, primary human liver cells are in short supply and display donor-to-donor variability in viability/quality. In contrast, human hepatocyte-like cells (HLCs) differentiated from induced pluripotent stem cells and embryonic stem cells are a near infinite cell resource that retains the patient/donor's genetic background; however, conventional protocols yield immature phenotypes. HLC maturation can be significantly improved using advanced techniques, such as protein micropatterning to precisely control cell-cell interactions, controlled sized spheroids, organoids with multiple cell types and layers, 3-dimensional bioprinting to spatially control cell populations, microfluidic devices for automated nutrient exchange and to induce liver zonation via soluble factor gradients, and synthetic biology to genetically modify the HLCs to accelerate and enhance maturation. Here, we present design features and characterization for representative advanced HLC maturation platforms and then discuss HLC use for modeling various liver diseases. Lastly, we discuss desirable advances to move this field forward. We anticipate that with continued advances in this space, pluripotent stem cell-derived liver models will provide human-relevant data much earlier in preclinical drug development and reduce animal usage, help elucidate liver disease mechanisms for the discovery of efficacious and safe therapeutics, and be useful as cell-based therapies for patients suffering from end-stage liver failure.
几种肝脏疾病（例如乙型/丙型肝炎病毒、酒精/非酒精性脂肪肝、疟疾、单基因疾病和药物性肝损伤）对全球的死亡率和发病率有重大影响。肝脏功能的物种特异性差异限制了动物在充分阐明/预测人类结果中的应用；因此，体外人类肝脏模型被用于基础和转化研究，以补充动物研究。然而，原代人肝细胞的供应有限，并且在活力/质量方面存在供体间的可变性。相比之下，由诱导多能干细胞和胚胎干细胞分化而来的人肝样细胞（HLC）是一种近乎无限的细胞资源，保留了患者/供体的遗传背景；然而，传统方案产生的表型不成熟。使用先进技术可以显著改善 HLC 的成熟度，例如蛋白质微图案化以精确控制细胞-细胞相互作用、控制大小的球体、具有多种细胞类型和层的类器官、用于空间控制细胞群体的 3 维生物打印、用于自动营养交换和通过可溶性因子梯度诱导肝区带形成的微流控装置，以及用于遗传修饰 HLC 以加速和增强成熟度的合成生物学。在这里，我们提出了代表性先进的 HLC 成熟平台的设计特点和表征，然后讨论了 HLC 在模拟各种肝脏疾病中的应用。最后，我们讨论了推动该领域发展的可取进展。我们预计，随着该领域的持续进步，多能干细胞衍生的肝脏模型将在临床前药物开发的早期提供更具人类相关性的数据，并减少动物的使用，有助于阐明肝脏疾病的机制，以发现有效和安全的治疗方法，并作为细胞疗法用于患有终末期肝功能衰竭的患者。
