Semi-automated Production of Hepatocyte Like Cells from Pluripotent Stem Cells.

Semi-automated Production of Hepatocyte Like Cells from Pluripotent Stem Cells.
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从多能干细胞半自动化生产肝细胞样细胞。

DOI:
10.3791/57995
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发表时间:
2018-07-27
期刊:
Journal of visualized experiments : JoVE
影响因子:
--
通讯作者:
Hay DC
Hay DC
中科院分区:
其他
文献类型:
--
作者:
Meseguer-Ripolles J;Lucendo-Villarin B;Wang Y;Hay DC

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人类多能干细胞代表了人类组织的可再生来源。我们的研究重点是从人类胚胎干细胞(hESC)和诱导多能干细胞(iPSC)中产生人类肝脏组织。目前的分化程序产生的人肝细胞样细胞(HLC)显示胎儿和成人性状的混合物。为了改善细胞表型,我们已经完全定义了我们的分化程序和细胞生态位,从而产生显示改善的基因表达和功能的细胞群。虽然这些研究标志着进步,但产生大量用于筛选的多孔板的能力受到劳动密集型程序和批次间差异的限制。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HLC。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光度计分析细胞表型。
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.
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