Wang Yan, Lee Jae-Ho, Shirahama Hitomi, Seo Jeongeun, Glenn Jeffrey S, Cho Nam-Joon
School of Materials Science and Engineering, Nanyang Technological University, 50 Nanyang Avenue 639798, Singapore.
Division of Gastroenterology and Hepatology, Department of Medicine, Stanford University School of Medicine, Alway Building, Room M211, 300 Pasteur Drive, Stanford, California 94305, United States.
ACS Biomater Sci Eng. 2016 Dec 12;2(12):2255-2265. doi: 10.1021/acsbiomaterials.6b00487. Epub 2016 Oct 28.
In this study, we constructed a microporous hydrogel scaffold with hexagonally packed interconnected cavities and extracellular matrix (ECM)-functionalized interior surface, and systematically investigated the hepatic differentiation of human adipose-derived mesenchymal stem cells (hAD-MSCs) under the influence of three key factors: three-dimensional (3D) geometry, ECM presence, and coculture with hepatocyte-derived cell line. Results confirmed that (i) hepatic differentiation of hAD-MSC is more efficient in a 3D microporous scaffold than in 2D monolayer culture; (ii) the presence of both ECM components (fibronectin and collagen-I) in the scaffold is superior to collagen-I only, highlighting the importance of fibronectin; and (iii) coculture with Huh-7.5 hepatocyte-derived cells promoted liver-specific functions of the hAD-MSC-derived hepatocytes. The optimized differentiation process only took 21 days to complete, a time length that is shorter or at least comparable to previous reports, and more importantly, yielded an albumin production more than 10-fold higher than conventional 2D culture. Our approach of optimizing hAD-MSC hepatic differentiation could provide a potential solution to the challenges such as hepatocyte transplantation or the establishment of human physiologically relevant liver models .
在本研究中，我们构建了一种具有六边形排列的相互连接腔室和细胞外基质（ECM）功能化内表面的微孔水凝胶支架，并系统地研究了在三维（3D）几何形状、ECM存在以及与肝细胞系共培养这三个关键因素影响下，人脂肪来源间充质干细胞（hAD-MSCs）的肝向分化。结果证实：（i）hAD-MSC在3D微孔支架中的肝向分化比在2D单层培养中更有效；（ii）支架中同时存在两种ECM成分（纤连蛋白和I型胶原）优于仅存在I型胶原，突出了纤连蛋白的重要性；（iii）与Huh-7.5肝细胞系共培养促进了hAD-MSC来源肝细胞的肝脏特异性功能。优化后的分化过程仅需21天即可完成，这一时间长度短于或至少与先前报道相当，更重要的是，其白蛋白产量比传统2D培养高出超过10倍。我们优化hAD-MSC肝向分化的方法可为肝细胞移植或建立与人类生理相关的肝脏模型等挑战提供潜在解决方案。
