Bio-Industry Unit, Technology Research Laboratory, Shimadzu Corporation, 3-9-4, Hikaridai, Seika-cho, Soraku-gun, Kyoto 619-0237, Japan.
Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6, Yamadaoka, Suita, Osaka 565-0871, Japan.
ACS Appl Bio Mater. 2021 Sep 20;4(9):7290-7299. doi: 10.1021/acsabm.1c00816. Epub 2021 Sep 8.
Various three-dimensional (3D) culture systems are available to provide more accurate in vivo mimicry than two-dimensional (2D) cultures. Synthetic and/or xeno-free biomaterials are desired, as they would provide lower batch-to-batch variability and high repeatability. Here, we introduce a 3D culture system using nanofibers composed of an amphiphilic polydepsipeptide-based polymer named HYDROX, which turns into 3D nanofibers after hydration. Our system produces a large amount of cell aggregates and requires only the seeding of a cell mixture. In addition, cells cultured with HYDROX can be collected with only a centrifugation procedure, and analytical assays can then be performed. Here, we applied HYDROX to hepatic differentiation from induced pluripotent stem cells. The cells cultured with HYDROX formed aggregates and HYDROX strongly promoted hepatic differentiation and maturation in terms of functions such as the positive ratio of alpha-1 antitrypsin, the production of albumin, the cytochrome P450 (CYP) 3A4 activity, and the low-density-lipoprotein uptake ability. In addition, primary human hepatocytes cultured with HYDROX showed significantly improved gene expression and activity. The viscoelasticity and stiffness of HYDROX can be modulated by varying the concentration of the synthetic polymer, thereby providing a suitable microenvironment for the differentiation of cells with various characteristics toward a target cell type. Our findings demonstrated that HYDROX is a promising biomaterial for 3D cultures in research fields ranging from basic cell research to drug discovery.
各种三维（3D）培养系统可提供比二维（2D）培养更准确的体内模拟，合成和/或无动物源的生物材料是理想的选择，因为它们可以提供更低的批次间变异性和更高的可重复性。在这里，我们引入了一种使用由名为 HYDROX 的两亲性聚肽基聚合物组成的纳米纤维的 3D 培养系统，该聚合物在水合后变成 3D 纳米纤维。我们的系统产生大量的细胞聚集体，只需要接种细胞混合物。此外，用 HYDROX 培养的细胞可以仅通过离心程序收集，然后可以进行分析测定。在这里，我们将 HYDROX 应用于诱导多能干细胞向肝系的分化。用 HYDROX 培养的细胞形成聚集体，并且 HYDROX 强烈促进了肝分化和成熟，表现在α-1 抗胰蛋白酶阳性率、白蛋白产生、细胞色素 P450（CYP）3A4 活性和低密度脂蛋白摄取能力等方面。此外，用 HYDROX 培养的原代人肝细胞显示出显著改善的基因表达和活性。通过改变合成聚合物的浓度可以调节 HYDROX 的粘弹性和硬度，从而为具有各种特性的细胞向目标细胞类型的分化提供合适的微环境。我们的研究结果表明，HYDROX 是一种很有前途的生物材料，可用于从基础细胞研究到药物发现的各种 3D 培养研究领域。
