Characterizing the effects of heparin gel stiffness on function of primary hepatocytes.
Characterizing the effects of heparin gel stiffness on function of primary hepatocytes.
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DOI：
10.1089/ten.tea.2012.0681
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发表时间：
2013-08
影响因子：
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通讯作者：
Jungmok You;Sua Park;Dong-Sik Shin;Dipali Patel;V. K. Raghunathan;Mihye Kim;C. Murphy;G. Tae
中科院分区：
文献类型：
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作者：
Jungmok You;Sua Park;Dong-Sik Shin;Dipali Patel;V. K. Raghunathan;Mihye Kim;C. Murphy;G. Tae
在肝脏中，肝细胞暴露于形成肝脏表型的大量刺激。当肝细胞在标准细胞培养皿中培养时，这种体内微环境丧失，使得在体外维持肝细胞功能具有挑战性。我们的文章集中在一个研究最少的诱导剂的肝表型-机械性能的基础基板。通过自由基介导的硫醇-烯偶联反应，在玻璃基底上形成由硫醇化肝素（Hep-SH）和双丙烯酸酯化聚乙二醇（PEG-DA）组成的凝胶层。通过改变前体溶液的浓度，基板刚度从10到110 kPa变化。ELISA分析显示，5天后，在较软的肝素凝胶上培养的肝细胞合成的白蛋白水平是在较硬的肝素凝胶上培养的肝细胞的5倍。免疫荧光染色的肝标志物，白蛋白和E-钙粘蛋白，证实了更软的凝胶促进更好地维持肝脏表型。我们的研究结果指出了基质机械性能对肝细胞功能的重要性。
In the liver, hepatocytes are exposed to a large array of stimuli that shape hepatic phenotype. This in vivo microenvironment is lost when hepatocytes are cultured in standard cell cultureware, making it challenging to maintain hepatocyte function in vitro. Our article focused on one of the least studied inducers of the hepatic phenotype-the mechanical properties of the underlying substrate. Gel layers comprised of thiolated heparin (Hep-SH) and diacrylated poly(ethylene glycol) (PEG-DA) were formed on glass substrates via a radical mediated thiol-ene coupling reaction. The substrate stiffness varied from 10 to 110 kPa by changing the concentration of the precursor solution. ELISA analysis revealed that after 5 days, hepatocytes cultured on a softer heparin gel were synthesizing five times higher levels of albumin compared to those on a stiffer heparin gel. Immunofluorescent staining for hepatic markers, albumin and E-cadherin, confirmed that softer gels promoted better maintenance of the hepatic phenotype. Our findings point to the importance of substrate mechanical properties on hepatocyte function.