Lucendo-Villarin Balta, Rashidi Hassan, Alhaque Sharmin, Fischer Lena, Meseguer-Ripolles Jose, Wang Yu, O'Farrelly Cliona, Themis Michael, Hay David C
MRC Centre for Regenerative Medicine, University of Edinburgh.
MRC Centre for Regenerative Medicine, University of Edinburgh; UCL Great Ormond Street Institute of Child Health, University College London.
J Vis Exp. 2019 Jul 20(149). doi: 10.3791/59965.
The development of renewable sources of liver tissue is required to improve cell-based modelling, and develop human tissue for transplantation. Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) represent promising sources of human liver spheres. We have developed a serum free and defined method of cellular differentiation to generate three-dimensional human liver spheres formed from human pluripotent stem cells. A potential limitation of the technology is the production of dense spheres with dead material inside. In order to circumvent this, we have employed agarose microwell technology at defined cell densities to control the size of the 3D spheres, preventing the generation of apoptotic and/or necrotic cores. Notably, the spheres generated by our approach display liver function and stable phenotype, representing a valuable resource for basic and applied scientific research. We believe that our approach could be used as a platform technology to develop further tissues to model and treat human disease and in the future may permit the generation of human tissue with complex tissue architecture.
为了改进基于细胞的建模，并开发用于移植的人体组织，需要开发可再生的肝脏组织来源。人类胚胎干细胞（hESCs）和人类诱导多能干细胞（hiPSCs）是有前景的人类肝球来源。我们已经开发出一种无血清且明确的细胞分化方法，以生成由人类多能干细胞形成的三维人类肝球。该技术的一个潜在限制是产生内部有死亡物质的致密球体。为了规避这一问题，我们采用琼脂糖微孔技术，在特定的细胞密度下控制3D球体的大小，防止凋亡和/或坏死核心的产生。值得注意的是，我们方法产生的球体具有肝功能和稳定的表型，是基础和应用科学研究的宝贵资源。我们相信，我们的方法可以用作平台技术，进一步开发用于建模和治疗人类疾病的组织，并且在未来可能允许生成具有复杂组织结构的人体组织。
