Generating size-controlled embryoid bodies using laser direct-write.
Generating size-controlled embryoid bodies using laser direct-write.
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DOI：
10.1088/1758-5082/6/2/025007
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发表时间：
2014-06
期刊：
影响因子：
9
通讯作者：
Corr DT
中科院分区：
文献类型：
--
作者：
Dias AD;Unser AM;Xie Y;Chrisey DB;Corr DT
胚胎干细胞（ESCs）具有自我更新和分化为任何特化细胞类型的潜力。体外分化ESCs的一种常用方法是通过胚状体（EBs），这是一种三维细胞聚集体，可以自发自组装，通常表达内胚层、外胚层和中胚层的标记物。先前的研究表明，EB大小和2D集落大小都会影响分化。我们假设我们可以通过细胞打印方法激光直写（LDW）来控制小鼠ESCs （mESCs）形成的EB的大小，以控制初始打印集落的大小和打印集落的局部细胞密度。在不同的印刷集落大小和印刷密度下印刷mESCs后，双向方差分析表明，印刷密度在3天后对EB直径有影响（p = 0.0002），而同一时间点印刷集落直径对EB直径没有影响（p = 0.74）。这两个因素之间没有显著的交互作用。Tukey的honest Significant Difference测试显示高密度集落形成了明显更大的EBs，这表明打印的mESCs与附近的细胞迅速聚集。因此，EBs可以通过控制印刷密度来设计到所需的尺寸，这将影响未来差异化研究的设计。在此，我们强调了LDW在规定的空间位置独立控制局部细胞密度和集落大小的能力，这可能导致更好的干细胞维持和定向分化。
Embryonic stem cells (ESCs) have the potential to self-renew and differentiate into any specialized cell type. One common method to differentiate ESCs in vitro is through embryoid bodies (EBs), 3D cellular aggregates that spontaneously self-assemble and generally express markers for the three germ layers, endoderm, ectoderm, and mesoderm. It has been previously shown that both EB size and 2D colony size each influence differentiation. We hypothesized that we could control the size of the EB formed by mouse ESCs (mESCs) by using a cell printing method, laser direct-write (LDW), to control both the size of the initial printed colony and the local cell density in printed colonies. After printing mESCs at various printed colony sizes and printing densities, two-way ANOVAs indicated that EB diameter was influenced by printing density after 3 days (p = 0.0002), while there was no effect of printed colony diameter on EB diameter at the same timepoint (p = 0.74). There was no significant interaction between these two factors. Tukey’s Honestly Significant Difference test showed that high-density colonies formed significantly larger EBs, suggesting that printed mESCs quickly aggregate with nearby cells. Thus, EBs can be engineered to a desired size by controlling printing density, which will influence the design of future differentiation studies. Herein, we highlight the capacity of LDW to control the local cell density and colony size independently, at prescribed spatial locations, potentially leading to better stem cell maintenance and directed differentiation.
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影响因子：
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