Meseguer-Ripolles Jose, Lucendo-Villarin Baltasar, Wang Yu, Hay David C
MRC Centre for Regenerative Medicine, University of Edinburgh.
MRC Centre for Regenerative Medicine, University of Edinburgh;
J Vis Exp. 2018 Jul 27(137):57995. doi: 10.3791/57995.
Human pluripotent stem cells represent a renewable source of human tissue. Our research is focused on generating human liver tissue from human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs). Current differentiation procedures generate human hepatocyte-like cells (HLCs) displaying a mixture of fetal and adult traits. To improve cell phenotype, we have fully defined our differentiation procedure and the cell niche, resulting in the generation of cell populations which display improved gene expression and function. While these studies mark progress, the ability to generate large quantities of multi well plates for screening has been limited by labour intensive procedures and batch to batch variation. To tackle this issue, we have developed a semi-automated platform to differentiate pluripotent stem cells into HLCs. Stem cell seeding and differentiation were performed using liquid handling and automatic pipetting systems in 96-well plate format. Following the differentiation, cell phenotype was analyzed using automated microscopy and a multi well luminometer.
人类多能干细胞代表了一种可再生的人体组织来源。我们的研究重点是从人类胚胎干细胞(hESCs)和诱导多能干细胞(iPSCs)中生成人类肝脏组织。目前的分化程序可生成具有胎儿和成人特征混合的人类肝细胞样细胞(HLCs)。为了改善细胞表型,我们已经完全确定了我们的分化程序和细胞微环境,从而产生了具有改善的基因表达和功能的细胞群体。虽然这些研究取得了进展,但由于劳动密集型程序和批次间差异,生成大量用于筛选的多孔板的能力受到了限制。为了解决这个问题,我们开发了一个半自动化平台,将多能干细胞分化为HLCs。使用液体处理和自动移液系统以96孔板形式进行干细胞接种和分化。分化后,使用自动显微镜和多孔发光计分析细胞表型。