Acta Biomaterialia ( IF 9.4 ) Pub Date : 2018-06-05 , DOI: 10.1016/j.actbio.2018.05.052 Elena A Silantyeva 1 , Wafaa Nasir 2 , Jacqueline Carpenter 2 , Olivia Manahan 1 , Matthew L Becker 3 , Rebecca K Willits 2
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Substrates for embryonic stem cell culture are typified by poorly defined xenogenic, whole proteins or cellular components that are difficult and expensive to generate, characterize, and recapitulate. Herein, the generation of well-defined scaffolds of Gly-Tyr-Ile-Gly-Ser-Arg (GYIGSR) peptide-functionalized poly(ε-caprolactone) (PCL) aligned nanofibers are used to accelerate the neural lineage commitment and differentiation of D3 mouse embryonic stem cells (mESCs). Gene expression trends and immunocytochemistry analysis were similar to laminin-coated glass, and indicated an earlier differentiation progression than D3 mESCs on laminin. Further, GYIGSR-functionalized nanofiber substrates yielded an increased gene expression of Sox1, a neural progenitor cell marker, and Tubb3, Cdh2, Syp, neuronal cell markers, at early time points. In addition, guidance of neurites was found to parallel the fiber direction. Herein, we demonstrate the fabrication of a well-defined, xeno-free functional nanofiber scaffold and demonstrates its use as a surrogate for xenogenic and complex matrixes currently used for the neural differentiation of stem cells ex vivo
Statement of Significance
In this paper, we report the use of GYIGSR-functionalized poly(ε-caprolactone) aligned nanofibers as a tool to accelerate the neural lineage commitment and differentiation of D3 mouse embryonic stem cells. The results indicate that functional nanofiber substrates promote faster differentiation than laminin coated substrates. The data suggest that aligned nanofibers and post-electrospinning surface modification with bioactive species can be combined to produce translationally relevant xeno-free substrates for stem cell therapy. Future development efforts are focused on additional bioactive species that are able to function as surrogates for other xenogenic factors found in differentiation media.
中文翻译：
对齐的 GYIGSR 功能化纳米纤维上加速小鼠胚胎干细胞的神经分化
胚胎干细胞培养的基质的典型特征是定义不明确的异种、完整蛋白质或细胞成分，这些成分的生成、表征和重演困难且昂贵。在此，生成明确的 Gly-Tyr-Ile-Gly-Ser-Arg (GYIGSR) 肽功能化聚(ε-己内酯) ( PCL) 排列纳米纤维支架，用于加速 D3 的神经谱系定型和分化小鼠胚胎干细胞（mESC）。基因表达趋势和免疫细胞化学分析与层粘连蛋白包被的玻璃相似，并且表明比层粘连蛋白上的 D3 mESC 更早的分化进程。此外，GYIGSR 功能化纳米纤维基底在早期时间点提高了Sox1（一种神经祖细胞标记物）和Tubb3、Cdh2、Syp （神经元细胞标记物）的基因表达。此外，发现神经突的引导平行于纤维方向。在此，我们展示了一种明确的、无异种的功能性纳米纤维支架的制造，并展示了其作为目前用于干细胞离体神经分化的异种和复杂基质的替代品的用途
重要性声明
在本文中，我们报告了使用 GYIGSR 功能化聚（ε-己内酯）排列纳米纤维作为加速 D3 小鼠胚胎干细胞的神经谱系定型和分化的工具。结果表明，功能性纳米纤维基底比层粘连蛋白涂层基底促进更快的分化。数据表明，排列好的纳米纤维和生物活性物质的静电纺丝后表面修饰可以结合起来，产生用于干细胞治疗的翻译相关的无异源基质。未来的开发工作重点是能够充当分化培养基中其他异种因子替代品的其他生物活性物种。
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