Optimizing human embryonic stem cells differentiation efficiency by screening size-tunable homogenous embryoid bodies
Optimizing human embryonic stem cells differentiation efficiency by screening size-tunable homogenous embryoid bodies
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DOI：
10.1016/j.biomaterials.2014.04.001
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发表时间：
2014-07-01
期刊：
影响因子：
14
通讯作者：
Lee, Sang-Hoon
中科院分区：
文献类型：
--
作者：
Moon, Sung-Hwan;Ju, Jongil;Lee, Sang-Hoon
人胚胎干细胞（hESC）通常通过在可溶性生长因子存在下形成称为胚状体（EB）的球形结构来诱导分化。 hEB 是通过悬浮小团 hESC 集落产生的；然而，所得的 hEB 是异质的，因为该方法缺乏控制单个 EB 中细胞数量的能力。这种异质性会影响影响分化的因素，例如细胞接触和可溶性因子的扩散，因此，每个 EB 的分化能力各不相同。在这里，我们制造了尺寸可调的凹形微孔来控制物理环境，从而调节单个 hESC 形成的 EB 的大小。指定数量的单个 hESC 被迫聚集并产生高保真度的均匀大小的 EB，并且使用不同直径的凹形微孔控制 EB 的大小。在生长因子不存在和存在的情况下，H9-和 CHA15-hESC 的分化模式均受到 EB 大小的影响。通过在不同 BMP4 浓度存在下筛选 EB 大小，实现了内皮细胞分化的两倍增加。由于每个 hESC 系都有独特的特征，本研究的结果表明，凹形微孔可用于筛选不同的 EB 大小和生长因子浓度，以优化每个 hESC 系的分化。 (C) 2014 Elsevier Ltd. 保留所有权利。
Human embryonic stem cells (hESCs) are generally induced to differentiate by forming spherical structures termed embryoid bodies (EBs) in the presence of soluble growth factors. hEBs are generated by suspending small clumps of hESC colonies; however, the resulting hEBs are heterogeneous because this method lacks the ability to control the number of cells in individual EBs. This heterogeneity affects factors that influence differentiation such as cell cell contact and the diffusion of soluble factors, and consequently, the differentiation capacity of each EB varies. Here, we fabricated size-tunable concave microwells to control the physical environment, thereby regulating the size of EBs formed from single hESCs. Defined numbers of single hESCs were forced to aggregate and generate uniformly sized EBs with high fidelity, and the size of the EBs was controlled using concave microwells of different diameters. Differentiation patterns in H9- and CHA15-hESCs were affected by EB size in both the absence and presence of growth factors. By screening EB size in the presence of various BMP4 concentrations, a twofold increase in endothelial cell differentiation was achieved. Because each hESC line has unique characteristics, the findings of this study demonstrate that concave microwells could be used to screen different EB sizes and growth factor concentrations to optimize differentiation for each hESC line. (C) 2014 Elsevier Ltd. All rights reserved.