Scalable topographies to support proliferation and Oct4 expression by human induced pluripotent stem cells.
Scalable topographies to support proliferation and Oct4 expression by human induced pluripotent stem cells.
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DOI：
10.1038/srep18948
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发表时间：
2016-01-13
影响因子：
4.6
通讯作者：
Watt FM
中科院分区：
文献类型：
--
作者：
Reimer A;Vasilevich A;Hulshof F;Viswanathan P;van Blitterswijk CA;de Boer J;Watt FM
地形特征可以调节细胞行为，包括细胞形态、增殖和分化。为了确定地形对人类诱导多能干细胞（iPSC）的影响，我们将细胞放置在包含1000多种不同特征的地形文库中，在缺乏动物产品（无xeno）的培养基中。利用高含量成像技术，我们测定了每种地形对细胞增殖和多能性标志物Oct4在播种后24 h表达的影响。维持Oct4表达的特征在24小时内也支持细胞增殖和细胞间粘附，4天后形成Oct4阳性和sox2阳性细胞的菌落。计算分析表明，小特征尺寸是决定多能性的最重要因素，其次是高波数和高特征密度。利用这些信息，我们正确地预测了库中任何给定的地形在24 h时是否支持多能态。这种方法不仅有助于设计最佳的人类iPSC扩增底物，而且还可能识别具有其他理想特征的地形，例如促进分化。
It is well established that topographical features modulate cell behaviour, including cell morphology, proliferation and differentiation. To define the effects of topography on human induced pluripotent stem cells (iPSC), we plated cells on a topographical library containing over 1000 different features in medium lacking animal products (xeno-free). Using high content imaging, we determined the effect of each topography on cell proliferation and expression of the pluripotency marker Oct4 24 h after seeding. Features that maintained Oct4 expression also supported proliferation and cell-cell adhesion at 24 h, and by 4 days colonies of Oct4-positive, Sox2-positive cells had formed. Computational analysis revealed that small feature size was the most important determinant of pluripotency, followed by high wave number and high feature density. Using this information we correctly predicted whether any given topography within our library would support the pluripotent state at 24 h. This approach not only facilitates the design of substrates for optimal human iPSC expansion, but also, potentially, identification of topographies with other desirable characteristics, such as promoting differentiation.