Tasnim Farah, Toh Yi-Chin, Qu Yinghua, Li Huan, Phan Derek, Narmada Balakrishnan C, Ananthanarayanan Abhishek, Mittal Nikhil, Meng Ryan Q, Yu Hanry
Institute of Bioengineering and Nanotechnology , #04-01, 31 Biopolis Way, The Nanos, Singapore 138669, Singapore.
Preclinical Development and Safety, Asia Pacific, Janssen Research & Development , 999 South Pudong Road, Shanghai, 200120, China.
Mol Pharm. 2016 Jun 6;13(6):1947-57. doi: 10.1021/acs.molpharmaceut.6b00119. Epub 2016 May 24.
Pluripotent stem cell derived hepatocyte-like cells (hPSC-HLCs) are an attractive alternative to primary human hepatocytes (PHHs) used in applications ranging from therapeutics to drug safety testing studies. It would be critical to improve and maintain mature hepatocyte functions of the hPSC-HLCs, especially for long-term studies. If 3D culture systems were to be used for such purposes, it would be important that the system can support formation and maintenance of optimal-sized spheroids for long periods of time, and can also be directly deployed in liver drug testing assays. We report the use of 3-dimensional (3D) cellulosic scaffold system for the culture of hPSC-HLCs. The scaffold has a macroporous network which helps to control the formation and maintenance of the spheroids for weeks. Our results show that culturing hPSC-HLCs in 3D cellulosic scaffolds increases functionality, as demonstrated by improved urea production and hepatic marker expression. In addition, hPSC-HLCs in the scaffolds exhibit a more mature phenotype, as shown by enhanced cytochrome P450 activity and induction. This enables the system to show a higher sensitivity to hepatotoxicants and a higher degree of similarity to PHHs when compared to conventional 2D systems. These results suggest that 3D cellulosic scaffolds are ideal for the long-term cultures needed to mature hPSC-HLCs. The mature hPSC-HLCs with improved cellular function can be continually maintained in the scaffolds and directly used for hepatotoxicity assays, making this system highly attractive for drug testing applications.
多能干细胞衍生的肝细胞样细胞（hPSC-HLCs）是用于从治疗到药物安全性测试研究等各种应用的原代人肝细胞（PHHs）的一种有吸引力的替代方案。改善和维持hPSC-HLCs的成熟肝细胞功能至关重要，特别是对于长期研究而言。如果将三维（3D）培养系统用于此类目的，那么该系统能够长时间支持形成和维持最佳大小的球体，并且还能够直接应用于肝脏药物测试分析，这一点很重要。我们报告了使用三维（3D）纤维素支架系统培养hPSC-HLCs。该支架具有大孔网络，有助于在数周内控制球体的形成和维持。我们的结果表明，在3D纤维素支架中培养hPSC-HLCs可提高其功能，尿素生成增加和肝脏标志物表达改善证明了这一点。此外，支架中的hPSC-HLCs表现出更成熟的表型，细胞色素P450活性增强和诱导作用表明了这一点。与传统的二维系统相比，这使得该系统对肝毒性物质表现出更高的敏感性，并且与PHHs具有更高的相似程度。这些结果表明，3D纤维素支架非常适合hPSC-HLCs成熟所需的长期培养。具有改善的细胞功能的成熟hPSC-HLCs可以在支架中持续维持，并直接用于肝毒性分析，使得该系统在药物测试应用中极具吸引力。
