Wang Yu, Alhaque Sharmin, Cameron Kate, Meseguer-Ripolles Jose, Lucendo-Villarin Baltasar, Rashidi Hassan, Hay David C
MRC Centre for Regenerative Medicine, University of Edinburgh.
MRC Centre for Regenerative Medicine, University of Edinburgh;
J Vis Exp. 2017 Mar 2(121):55355. doi: 10.3791/55355.
Human pluripotent stem cells (hPSCs) possess great value for biomedical research. hPSCs can be scaled and differentiated to all cell types found in the human body. The differentiation of hPSCs to human hepatocyte-like cells (HLCs) has been extensively studied, and efficient differentiation protocols have been established. The combination of extracellular matrix and biological stimuli, including growth factors, cytokines, and small molecules, have made it possible to generate HLCs that resemble primary human hepatocytes. However, the majority of procedures still employ undefined components, giving rise to batch-to-batch variation. This serves as a significant barrier to the application of the technology. To tackle this issue, we developed a defined system for hepatocyte differentiation using human recombinant laminins as extracellular matrices in combination with a serum-free differentiation process. Highly efficient hepatocyte specification was achieved, with demonstrated improvements in both HLC function and phenotype. Importantly, this system is easy to scale up using research and GMP-grade hPSC lines promising advances in cell-based modelling and therapies.
人多能干细胞（hPSCs）在生物医学研究中具有巨大价值。hPSCs可以扩增并分化为人体中发现的所有细胞类型。hPSCs向人肝细胞样细胞（HLCs）的分化已得到广泛研究，并已建立了有效的分化方案。细胞外基质与生物刺激因素（包括生长因子、细胞因子和小分子）的结合，使得生成类似于原代人肝细胞的HLCs成为可能。然而，大多数方法仍使用成分不明确的物质，导致批次间存在差异。这成为该技术应用的重大障碍。为解决这一问题，我们开发了一种明确的肝细胞分化系统，使用人重组层粘连蛋白作为细胞外基质，并结合无血清分化过程。实现了高效的肝细胞定向分化，HLC的功能和表型均有明显改善。重要的是，该系统易于使用研究级和GMP级hPSC系进行扩大规模，有望在基于细胞的建模和治疗方面取得进展。
