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丝纳米纤维电纺支架增强了从睾丸中分离得到的胚胎干细胞样细胞向成熟神经元的分化。

Silk nanofibrous electrospun scaffold enhances differentiation of embryonic stem like cells derived from testis in to mature neuron.

机构信息

Immunogenetic Research Center, Department of Anatomy & Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences, P.O. Box, Sari, 48471-91971, Iran.

Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.

出版信息

J Biomed Mater Res A. 2018 Oct;106(10):2662-2669. doi: 10.1002/jbm.a.36463.

Abstract

The scaffolds accompanied with stem cells have great potential for applications in neural tissue engineering. Fabrication of nanofibrous scaffold similar to extracellular matrix is one of the applicable methods in neural tissue regeneration. The aim of this study was the fabrication of a silk nanofibrous scaffold as a microenvironment for neural guiding differentiation of embryonic stem like cells (ES Like cells) derived from testis toward neuron-like cells. ES Like derived from culturing of testicular cells in vitro, were seeded on silk scaffolds and induced to neuronal phenotype using 4-/4± RA technique following culturing the cells in the neurobasal medium supplemented with 20 ng/mL bFGF,10 ng/mL EGF, B27, and N2 for 8-12 days. The neural differentiation was confirmed via the evaluation of specific neural markers; Nestin, NF68, MAP2 and β tubulin using immunocytochemistry and real-time polymerase chain reaction. Our results showed that silk scaffold support the attachment and proliferation of ES Like cells. The expression of Nestin, NF68, Map2, and ß tubulin markers were higher in cells grown on silk scaffold in compare to monolayer group. This study suggests electrospun silk nanofibrous scaffold as an appropriate substrate for neural induction of stem cells that is applicable for repairmen of damaged neural tissues. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2662-2669, 2018.

摘要

支架与干细胞联合具有在神经组织工程中应用的巨大潜力。 制造类似于细胞外基质的纳米纤维支架是神经组织再生的一种可行方法。本研究的目的是制造丝纳米纤维支架作为微环境,用于胚胎干细胞样细胞(ES 样细胞)从睾丸向神经元样细胞的神经导向分化。 ES 样细胞通过体外培养睾丸细胞获得,接种在丝支架上,并使用 4-/4±RA 技术在含有 20ng/mL bFGF、10ng/mL EGF、B27 和 N2 的神经基础培养基中培养 8-12 天,诱导其向神经元表型分化。通过免疫细胞化学和实时聚合酶链反应评估特定的神经标记物(巢蛋白、NF68、MAP2 和 β 微管蛋白)来确认神经分化。我们的结果表明,丝支架支持 ES 样细胞的附着和增殖。与单层组相比,在丝支架上生长的细胞中巢蛋白、NF68、Map2 和β微管蛋白标记物的表达更高。这项研究表明,静电纺丝纳米纤维支架是干细胞神经诱导的合适基质,适用于受损神经组织的修复。 © 2018 威利父子公司。J 生物医学材料研究部分 A:106A:2662-2669,2018。

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