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General Study and Gene Expression Profiling of Endotheliocytes Cultivated on Electrospun Materials

机译:电纺材料上培养的内皮细胞的一般研究和基因表达谱

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摘要

Endothelization of the luminal surface of vascular grafts is required for their long-term functioning. Here, we have cultivated human endothelial cells (HUVEC) on different 3D matrices to assess cell proliferation, gene expression and select the best substrate for endothelization. 3D matrices were produced by electrospinning from solutions of poly( , -lactide-co-glycolide) (PLGA), polycaprolactone (PCL), and blends of PCL with gelatin (Gl) in hexafluoroisopropanol. Structure and surface properties of 3D matrices were characterized by SEM, AFM, and sessile drop analysis. Cell adhesion, viability, and proliferation were studied by SEM, Alamar Blue staining, and 5-ethynyl-2’-deoxyuridine (EdU) assay. Gene expression profiling was done on an Illumina HiSeq 2500 platform. Obtained data indicated that 3D matrices produced from PCL with Gl and treated with glutaraldehyde provide the most suitable support for HUVEC adhesion and proliferation. Transcriptome sequencing has demonstrated a minimal difference of gene expression profile in HUVEC cultivated on the surface of these matrices as compared to tissue culture plastic, thus confirming these matrices as the best support for endothelization.
机译:血管移植物的腔表面需要内皮化才能长期发挥作用。在这里,我们在不同的3D基质上培养了人内皮细胞(HUVEC),以评估细胞增殖,基因表达并选择用于内皮化的最佳底物。通过静电纺丝从聚(-丙交酯-共-乙交酯)(PLGA),聚己内酯(PCL)以及PCL与明胶(GI)在六氟异丙醇中的混合物中制备3D基质。 3D矩阵的结构和表面特性通过SEM,AFM和固着分析进行了表征。通过SEM,Alamar Blue染色和5-乙炔基-2′-脱氧尿苷(EdU)分析研究了细胞粘附,活力和增殖。基因表达谱分析是在Illumina HiSeq 2500平台上进行的。所获得的数据表明,由PCL与G1产生并经戊二醛处理的3D基质为HUVEC粘附和增殖提供了最合适的支持。与组织培养塑料相比,转录组测序已证明在这些基质表面上培养的HUVEC中基因表达谱的差异最小,从而证实了这些基质是内皮化的最佳支持。

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