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Micro-scaled topographies direct differentiation of human epidermal stem cells.
Acta Biomaterialia ( IF 9.7 ) Pub Date : 2018-12-05 , DOI: 10.1016/j.actbio.2018.12.003
Sebastiaan Zijl 1 , Aliaksei S Vasilevich 2 , Priyalakshmi Viswanathan 1 , Ayelen Luna Helling 1 , Nick R M Beijer 2 , Gernot Walko 3 , Ciro Chiappini 4 , Jan de Boer 5 , Fiona M Watt 1
Affiliation  

Human epidermal stem cells initiate terminal differentiation when spreading is restricted on ECM-coated micropatterned islands, soft hydrogels or hydrogel-nanoparticle composites with high nanoparticle spacing. The effect of substrate topography, however, is incompletely understood. To explore this, primary human keratinocytes enriched for stem cells were seeded on a topographical library with over 2000 different topographies in the micrometre range. Twenty-four hours later the proportion of cells expressing the differentiation marker transglutaminase-1 was determined by high content imaging. As predicted, topographies that prevented spreading promoted differentiation. However, we also identified topographies that supported differentiation of highly spread cells. Topographies supporting differentiation of spread cells were more irregular than those supporting differentiation of round cells. Low topography coverage promoted differentiation of spread cells, whereas high coverage promoted differentiation of round cells. Based on these observations we fabricated a topography in 6-well plate format that supported differentiation of spread cells, enabling us to examine cell responses at higher resolution. We found that differentiated spread cells did not assemble significant numbers of hemidesmosomes, focal adhesions, adherens junctions, desmosomes or tight junctions. They did, however, organise the actin cytoskeleton in response to the topographies. Rho kinase inhibition and blebbistatin treatment blocked the differentiation of spread cells, whereas SRF inhibition did not. These observations suggest a potential role for actin polymerization and actomyosin contraction in the topography-induced differentiation of spread cells. STATEMENT OF SIGNIFICANCE: The epidermis is the outer covering of the skin. It is formed by layers of cells called keratinocytes. The basal cell layer contains stem cells, which divide to replace cells in the outermost layers that are lost through a process known as differentiation. In this manuscript we have developed surfaces that promote the differentiation of epidermal stem cells in order to understand the signals that control differentiation. The experimental tools we have developed have the potential to help us to devise new treatments that control diseases such as psoriasis and eczema in which epidermal stem cell proliferation and differentiation are disturbed.

中文翻译:

微型拓扑结构直接分化人类​​表皮干细胞。

当扩散受限于 ECM 涂层的微图案岛、软水凝胶或具有高纳米颗粒间距的水凝胶-纳米颗粒复合材料时,人表皮干细胞会启动终末分化。然而,基板形貌的影响还不完全清楚。为了探索这一点,将富含干细胞的原代人角质形成细胞接种在一个地形库中,该库具有超过 2000 种微米范围内的不同地形图。二十四小时后,通过高内涵成像确定表达分化标志物转谷氨酰胺酶-1 的细胞比例。正如预测的那样,阻止传播的地形促进了分化。然而,我们还确定了支持高度扩散细胞分化的拓扑结构。支持扩散细胞分化的拓扑结构比支持圆形细胞分化的拓扑结构更不规则。低地形覆盖促进扩散细胞的分化,而高覆盖促进圆形细胞的分化。基于这些观察,我们制作了支持扩散细胞分化的 6 孔板格式的地形图,使我们能够以更高分辨率检查细胞反应。我们发现分化的扩散细胞没有组装大量的半桥粒、粘着斑、粘附连接、桥粒或紧密连接。然而,他们确实根据地形组织了肌动蛋白细胞骨架。Rho 激酶抑制和 blebbistatin 治疗阻止了扩散细胞的分化,而 SRF 抑制则没有。这些观察表明肌动蛋白聚合和肌动球蛋白收缩在地形诱导的扩散细胞分化中的潜在作用。意义声明:表皮是皮肤的外层覆盖物。它由称为角质形成细胞的细胞层形成。基底细胞层包含干细胞,它们分裂以替换最外层中通过称为分化的过程丢失的细胞。在这份手稿中,我们开发了促进表皮干细胞分化的表面,以了解控制分化的信号。我们开发的实验工具有可能帮助我们设计出新的治疗方法来控制牛皮癣和湿疹等表皮干细胞增殖和分化受到干扰的疾病。
更新日期:2018-12-06
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