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生物活性分子修饰的表面增强了间充质干细胞的黏附与增殖。

Bio-active molecules modified surfaces enhanced mesenchymal stem cell adhesion and proliferation.

作者信息

Mobasseri Rezvan, Tian Lingling, Soleimani Masoud, Ramakrishna Seeram, Naderi-Manesh Hossein

机构信息

Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran; Center for Nanofibers & Nanotechnology, Department of Mechanical Engineering, National University of Singapore, 117576, Singapore.

Center for Nanofibers & Nanotechnology, Department of Mechanical Engineering, National University of Singapore, 117576, Singapore.

出版信息

Biochem Biophys Res Commun. 2017 Jan 29;483(1):312-317. doi: 10.1016/j.bbrc.2016.12.146. Epub 2016 Dec 23.

Abstract

Surface modification of the substrate as a component of in vitro cell culture and tissue engineering, using bio-active molecules including extracellular matrix (ECM) proteins or peptides derived ECM proteins can modulate the surface properties and thereby induce the desired signaling pathways in cells. The aim of this study was to evaluate the behavior of human bone marrow mesenchymal stem cells (hBM-MSCs) on glass substrates modified with fibronectin (Fn), collagen (Coll), RGD peptides (RGD) and designed peptide (R-pept) as bio-active molecules. The glass coverslips were coated with fibronectin, collagen, RGD peptide and R-peptide. Bone marrow mesenchymal stem cells were cultured on different substrates and the adhesion behavior in early incubation times was investigated using scanning electron microscopy (SEM) and confocal microscopy. The MTT assay was performed to evaluate the effect of different bio-active molecules on MSCs proliferation rate during 24 and 72 h. Formation of filopodia and focal adhesion (FA) complexes, two steps of cell adhesion process, were observed in MSCs cultured on bio-active molecules modified coverslips, specifically in Fn coated and R-pept coated groups. SEM image showed well adhesion pattern for MSCs cultured on Fn and R-pept after 2 h incubation, while the shape of cells cultured on Coll and RGD substrates indicated that they might experience stress condition in early hours of culture. Investigation of adhesion behavior, as well as proliferation pattern, suggests R-peptide as a promising bio-active molecule to be used for surface modification of substrate in supporting and inducing cell adhesion and proliferation.

摘要

作为体外细胞培养和组织工程的一个组成部分,对基质进行表面修饰,使用包括细胞外基质(ECM)蛋白或源自ECM蛋白的肽在内的生物活性分子,可以调节表面性质,从而在细胞中诱导所需的信号通路。本研究的目的是评估人骨髓间充质干细胞(hBM-MSCs)在经纤连蛋白(Fn)、胶原蛋白(Coll)、RGD肽(RGD)和设计肽(R-pept)修饰的玻璃基质上作为生物活性分子的行为。玻璃盖玻片用纤连蛋白、胶原蛋白、RGD肽和R肽进行包被。将骨髓间充质干细胞培养在不同的基质上,并使用扫描电子显微镜(SEM)和共聚焦显微镜研究早期孵育时间的黏附行为。进行MTT试验以评估不同生物活性分子在24小时和72小时内对间充质干细胞增殖率的影响。在生物活性分子修饰的盖玻片上培养的间充质干细胞中观察到丝状伪足和粘着斑(FA)复合物的形成,这是细胞黏附过程的两个步骤,特别是在Fn包被组和R-pept包被组中。SEM图像显示,孵育2小时后,在Fn和R-pept上培养的间充质干细胞具有良好的黏附模式,而在Coll和RGD基质上培养的细胞形状表明它们在培养早期可能经历应激状态。对黏附行为以及增殖模式的研究表明,R肽是一种有前途的生物活性分子,可用于基质的表面修饰,以支持和诱导细胞黏附和增殖。

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