Liver Regeneration Using Cultured Liver Bud
Liver Regeneration Using Cultured Liver Bud
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DOI：
10.1007/978-1-4939-6949-4_15
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发表时间：
2017-01-01
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影响因子：
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通讯作者：
Taniguchi, Hideki
中科院分区：
文献类型：
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作者：
Sekine, Keisuke;Takebe, Takanori;Taniguchi, Hideki
在这里，我们描述了一种在体外从人多能干细胞中培养三维（3D）肝芽样组织的方案。该方法主要由两部分组成：(1)人多能干细胞在二维培养中向肝内胚层（HE）分化；(2)多能干细胞与内皮细胞和间充质细胞共培养。首先，将iPSCs分化为最终内胚层（DE）细胞，将DE细胞分化为HE细胞，然后将其与内皮细胞和间充质细胞在基质涂覆塑料板或微纹板上共培养。细胞迅速凝聚形成三维组织团块。我们将这些iPSC肝芽命名为iPSC- lbs，因为它们在基因表达、细胞增殖和细胞比例方面与发育中的肝芽相似。这种肝芽培养系统为未来的临床应用、药物开发以及作为研究人类发育的工具提供了一种新的方法。
Here, we describe a protocol to develop a three-dimensional (3D) liver bud-like tissue from human iPSCs in vitro. This method mainly consists of two parts: (1) hepatic endoderm (HE) differentiation from human iPSCs in 2D culture and (2) co-culturing iPSC-HE with endothelial and mesenchymal cells. First, iPSCs were differentiated into definitive endoderm (DE) cells, and the DE cells were differentiated into HE cells, which were then co-cultured with endothelial cells and mesenchymal cells on Matrigel-coated plastic plates or micropattern plates. The cells rapidly condensed to generate 3D tissue masses. We named these iPSC liver buds (iPSC-LBs) because they resemble the developing liver bud from the perspective of gene expression, cell proliferation, and cell proportion. This liver bud culture system provides a novel approach for future clinical applications, for drug development, and as a tool for studying human development.