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首页> 外文期刊>Journal of tissue engineering and regenerative medicine >Hepatic differentiation of human embryonic stem cells on growth factor-containing surfaces.
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Hepatic differentiation of human embryonic stem cells on growth factor-containing surfaces.

机译：人胚胎干细胞在含生长因子的表面上的肝分化。

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Embryonic stem cells (ESCs) hold considerable promise in tissue engineering and regenerative medicine as a source of tissue-specific cells. Hepatocytes derived from ESCs will be useful for therapies, bioartificial liver assistance devices and drug discovery. In traditional stem cell cultivation/differentiation experiments, growth factors (GFs) are added in soluble form in order to provide signals for tissue-specific differentiation. In contrast, we investigated differentiation of hESCs cultured on top of GFs. In these experiments, glass substrates were imprinted with a mixture of ECM and GF molecules to form 500?μm diameter spots. hESCs were cultured onto these GF-containing ECM spots for up to 12?days to induce differentiation towards the hepatic lineage. The dynamics of differentiation were examined by quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) and immunocytochemistry. Stem cells cultured on GF-containing surfaces stained positive for the endoderm markers SOX17 and FOXA2, as well as early liver markers such as α-fetoprotein and albumin. qRT-PCR confirmed that pluripotency, endoderm and liver gene expression of hESCs cultured on GF-containing surfaces was consistent with hepatic differentiation. In comparison, hESCs cultured on ECM spots without GFs showed less pronounced loss of pluripotency and lower levels of liver gene expression. In summary, our study demonstrates that hESCs receive differentiation-inducing signals from GF-containing surfaces and may be pushed along the hepatic lineage when cultured on such surfaces. Compared to traditional approaches, the advantages of GF immobilization include reduction in the cost of experiments, stronger and longer stimulation and the possibility of screening GF-stem cell interactions in a multiplexed manner. Copyright ? 2012 John Wiley & Sons, Ltd.
机译：胚胎干细胞（ESC）作为组织特异性细胞的来源在组织工程和再生医学中具有广阔的前景。 ESC衍生的肝细胞将用于治疗，生物人工肝辅助设备和药物发现。在传统的干细胞培养/分化实验中，以可溶性形式添加生长因子（GFs），以提供组织特异性分化的信号。相反，我们研究了在GFs上培养的hESC的分化。在这些实验中，玻璃基板上印有ECM和GF分子的混合物，形成直径为500μm的斑点。将hESCs培养到这些含有GF的ECM点上长达12天，以诱导向肝谱系的分化。通过定量逆转录酶聚合酶链反应（qRT-PCR）和免疫细胞化学检查了分化的动力学。培养在含GF的表面上的干细胞对内胚层标记SOX17和FOXA2以及早期肝标记（例如甲胎蛋白和白蛋白）染色呈阳性。 qRT-PCR证实，在含GF的表面上培养的hESC的多能性，内胚层和肝基因表达与肝分化一致。相比之下，在没有GFs的ECM点上培养的hESCs表现出的多能性丧失不明显，肝脏基因表达水平较低。总而言之，我们的研究表明，hESCs从含有GF的表面接收诱导分化的信号，并且在此类表面上培养时可能沿肝谱系被推动。与传统方法相比，固定化GF的优势包括降低实验成本，更强和更长的刺激以及以多重方式筛选GF-干细胞相互作用的可能性。版权？ 2012年John Wiley＆Sons，Ltd.

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