Nitric oxide releasing hydrogel promotes endothelial differentiation of mouse embryonic stem cells
Nitric oxide releasing hydrogel promotes endothelial differentiation of mouse embryonic stem cells
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DOI：
10.1016/j.actbio.2017.08.037
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发表时间：
2017-11-01
影响因子：
9.7
通讯作者：
Li, Zongjin
中科院分区：
文献类型：
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作者：
Nie, Yan;Zhang, Kaiyue;Li, Zongjin
内皮细胞移植在治疗各种缺血性疾病方面具有广阔的前景。然而，在临床应用中，基于EC的治疗的主要挑战是提供高质量和足够量的细胞。在这项研究中，我们开发了一个简单而有效的系统，直接内皮分化的小鼠胚胎干细胞（ESCs）使用可控的壳聚糖一氧化氮（NO）释放水凝胶（CS-NO）。将ESCs接种于水凝胶培养系统上，并测量分化标志物的表达。结果发现，在控制NO释放的环境下，早期内皮细胞标志物Flk-1和成熟内皮细胞标志物VE-cadherin的表达明显增加。此外，Flk-1上调伴随着磷酸肌醇-3激酶（PI 3 K）/Akt信号传导的激活。我们还发现，在PI 3 K抑制剂（LY 294002）的存在下，ESCs的内皮定型被废除，表明Akt磷酸化在ESCs的内皮分化中的重要性。有趣的是，在没有NO的情况下，单独使用AICT激活剂（SC-79）激活Akt磷酸化并不能显著促进ESCs的内皮分化，这表明NO和Akt磷酸化在驱动ESCs的内皮命运特化中存在相互依赖的关系。两者合计，我们表明，NO释放在一个连续的和控制的方式是一个简单而有效的方法来指导内皮细胞分化的ESCs，而不添加growthfactors.Statement的SignificanceFastening数据继续表明，人工干细胞龛不仅作为一个物理支持支架干细胞增殖，但也作为一个新的平台，指导干细胞分化。由于缺乏合适的微环境在体外产生治疗性内皮细胞（EC），用于移植的EC的来源是基于EC的治疗的临床应用的主要限制。本研究建立了一种无饲养层细胞的二维培养体系，通过使用一氧化氮（NO）控释水凝胶（CS-NO）促进胚胎干细胞（ESCs）向内皮细胞分化。值得注意的是，从水凝胶释放的NO可以选择性地上调Flk-1（早期ECs标志物）和VE-钙粘蛋白（成熟ECs标志物），这在ESCs的内皮分化中是至关重要的。综上所述，本研究提出了一种简单有效的方法来指导ESCs的内皮分化，而无需额外的生长因子。(C)2017 Acta Materialia Inc.由爱思唯尔有限公司出版。保留所有权利。
Transplantation of endothelial cells (ECs) holds great promise for treating various kinds of ischemic diseases. However, the major challenge in ECs-based therapy in clinical applications is to provide high quality and enough amounts of cells. In this study, we developed a simple and efficient system to direct endothelial differentiation of mouse embryonic stem cells (ESCs) using a controllable chitosan nitric oxide (NO)-releasing hydrogel (CS-NO). ESCs were plated onto the hydrogel culture system, and the expressions of differentiation markers were measured. We found that the expression of Flk-1 (early ECs marker) and VE-cadherin (mature ECs marker) increased obviously under the controlled NO releasing environment. Moreover, the Flk-1 upregulation was accompanied by the activation of the phosphoinositide-3 kinase (PI3K)/Akt signaling. We also found that in the presence of the PI3K inhibitor (LY294002), the endothelial commitment of ESCs was abolished, indicating the importance of Akt phosphorylation in the endothelial differentiation of ESCs. Interestingly, in the absence of NO, the activation of Akt phosphorylation alone by using AICT activator (SC-79) did not profoundly promote the endothelial differentiation of ESCs, suggesting an interdependent relationship-between NO and the Akt phosphorylation in driving endothelial fate specification of ESCs. Taken together, we demonstrated that NO releasing in a continuous and controlled manner is a simple and efficient method for directing the endothelial differentiation of ESCs without adding growth factors.Statement of SignificanceFascinating data continues to show that artificial stem cell niche not only serve as a physical supporting scaffold for stem cells proliferation, but also as a novel platform for directing stem cell differentiation. Because of the lack of proper microenvironment for generating therapeutic endothelial cells (ECs) in vitro, the source of ECs for transplantation is the major limitation in ECs-based therapy to clinical applications. The current study established a feeder cell-free, 2-dimensional culture system for promoting the differentiation processes of embryonic stem cells (ESCs) committed to the endothelial lineage via using a nitric oxide (NO) controlled releasing hydrogel (CS-NO). Notably, the NO releasing from the hydrogel could selectively up-regulate Flk-1 (early ECs marker) and VE-cadherin (mature ECs marker) in the absence of growth factors, which was of crucial importance in the endothelial differentiation of ESCs. In summary, the current study proposes a simple and efficient method for directing the endothelial differentiation of ESCs without extra growth factors. (C) 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.