Oxygen Plasma Substrate and Specific Nanopattern Promote Early Differentiation of HepaRG Progenitors.
Oxygen Plasma Substrate and Specific Nanopattern Promote Early Differentiation of HepaRG Progenitors.
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氧等离子体底物和特定纳米图案促进 HepaRG 祖细胞的早期分化。
DOI：
10.1089/ten.tea.2019.0241
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发表时间：
2020
影响因子：
--
通讯作者：
Morgan K
中科院分区：
文献类型：
--
作者：
Morgan K
完全分化的HepaRG™细胞是毒理学和药物试验中体外研究的首选肝细胞系。它们来源于分化为肝细胞样和胆管细胞样细胞共培养物的成肝细胞样祖细胞（HepaRG-P）。这个过程需要2周的增殖，然后使用二甲基亚砜（DMSO）进行2周的分化，这可能是耗时且昂贵的。确定一种加速HepaRG-Ps向成熟谱系发展的方法将节省时间和金钱。在不存在DMSO的情况下做到这一点的能力将消除因DMSO诱导CYP途径而导致毒理学结果混淆的可能性。研究表明，组织培养基质在细胞系的发育和成熟过程中起着重要作用，而这一点对于祖细胞尤为重要，因为祖细胞仍具有一定的可塑性。氧等离子体处理被广泛用于修饰细胞培养基质。还有证据表明，图案化而不是平面表面对增殖和分化有积极影响。在这项研究中，我们比较了标准组织培养塑料（TCP）、氧等离子体涂层（OPC）和纳米图案化基底（OPC）对HepaRG-P细胞早期分化和功能的影响。由于OPC是OPC，我们最初比较了TCP和OPC的影响，以便使用OPC作为对照来评估图案化是否进一步增强早期分化和功能。结果表明，与TCP相比，在OPC底物上生长的HepaRG-P表现出更早的分化、增殖和功能。在OPC上培养HepaRG-P并添加β-内酰胺酶没有带来任何额外的优势。总之，OPC表面出现，以提高肝分化和功能，并可以取代传统的方法分化HepaRG-P细胞到完全分化和功能HepaRGs早于标准methods.Impact statementWe显示显着早期分化和功能的HepaRG祖细胞时，生长在二甲基亚砜无培养基上的氧等离子体基质与标准组织培养塑料。进一步的研究表明，氧等离子体基底的纳米图案化与平滑氧等离子体相比没有任何额外的优势，尽管一种图案（DSQ 120）显示出相当的早期分化和功能。
Fully differentiated HepaRG™ cells are the hepatic cell line of choice forin vitrostudy in toxicology and drug trials. They are derived from a hepatoblast-like progenitor (HepaRG-P) that differentiates into a coculture of hepatocyte-like and cholangiocyte-like cells. This process that requires 2 weeks of proliferation followed by 2 weeks of differentiation using dimethyl sulfoxide (DMSO) can be time consuming and costly. Identifying a method to accelerate HepaRG-Ps toward a mature lineage would save both time and money. The ability to do this in the absence of DMSO would remove the possibility of confounding toxicology results caused by DMSO induction of CYP pathways. It has been shown that tissue culture substrates play an important role in the development and maturity of a cell line, and this is particularly important for progenitor cells, which retain some form of plasticity.Oxygen plasma treatment is used extensively to modify cell culture substrates. There is also evidence that patterned rather than planar surfaces have a positive effect on proliferation and differentiation. In this study, we compared the effect of standard tissue culture plastic (TCP), oxygen plasma coated (OPC), and nanopatterned substrates (NPS) on early differentiation and function of HepaRG-P cells.Since NPS were OPC we initially compared the effect of TCP and OPC to enable comparison between all three culture surfaces using OPC as control to asses if patterning further enhanced early differentiation and functionality. The results show that HepaRG-P's grown on OPC substrate exhibited earlier differentiation, proliferation, and function compared with TCP. Culturing HepaRG-P's on OPC with the addition of NPS did not confer any additional advantage. In conclusion, OPC surface appeared to enhance hepatic differentiation and functionality and could replace traditional methods of differentiating HepaRG-P cells into fully differentiated and functional HepaRGs earlier than standard methods.Impact statementWe show significantly earlier differentiation and function of HepaRG progenitor cells when grown in dimethyl sulfoxide-free medium on oxygen plasma substrates versus standard tissue culture plastic. Further investigation showed that nanopatterning of oxygen plasma substrates did not confer any additional advantage over smooth oxygen plasma, although one pattern (DSQ120) showed comparable early differentiation and function.