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一种用于增强三维支架中细胞生长的有效装置和方法:静态和动态条件下细胞接种与增殖的研究

An effective device and method for enhanced cell growth in 3D scaffolds: Investigation of cell seeding and proliferation under static and dynamic conditions.

作者信息

Theodoridis Konstantinos, Aggelidou Eleni, Manthou Maria-Eleni, Keklikoglou Kleoniki, Tsimponis Antonios, Demiri Efterpi, Bakopoulou Athina, Mihailidis Athanasios, Kritis Aristeidis

机构信息

Department of Physiology and Pharmacology, School of Medicine, Faculty of Health Sciences, Aristotle University of Thessaloniki (A.U.Th), Thessaloniki, Greece; cGMP Regenerative Medicine Facility, Department of Physiology and Pharmacology, School of Medicine, Faculty of Health Sciences, Aristotle University of Thessaloniki (A.U.Th), Thessaloniki, Greece.

Department of Physiology and Pharmacology, School of Medicine, Faculty of Health Sciences, Aristotle University of Thessaloniki (A.U.Th), Thessaloniki, Greece; cGMP Regenerative Medicine Facility, Department of Physiology and Pharmacology, School of Medicine, Faculty of Health Sciences, Aristotle University of Thessaloniki (A.U.Th), Thessaloniki, Greece.

出版信息

Mater Sci Eng C Mater Biol Appl. 2020 Sep;114:111060. doi: 10.1016/j.msec.2020.111060. Epub 2020 May 11.

Abstract

Cell adhesion on 3D-scaffolds is a challenging task to succeed high cell densities and even cell distribution. We aimed to design a 3D-cell Culture Device (3D-CD) for static seeding and cultivation, to be used with any kind of scaffold, limiting cell loss and facilitating nutrient supply. 3D printing technology was used for both scaffold and device fabrication. Apart from testing the device, the purpose of this study was to assess and compare static and dynamic seeding and cultivation methods, of wet and dry scaffolds, under normoxic and hypoxic conditions and their effects on parameters such as cell seeding efficiency, cell distribution and cell proliferation. Human adipose tissue was harvested and cultured in 3D-printed poly(epsilon-caprolactone) scaffolds. Micro-CT scans were performed and projection images were reconstructed into cross section images. We created 3D images to visualize cell distribution and orientation inside the scaffolds. The group of prewetted scaffolds was the most favorable to cell attachment. The 3D-cell Culture Device (3D-CD) enhanced cell seeding efficiency with almost no cell loss. We suggest that the most favorable outcome can be produced with static seeding in the device for 24 h, followed either by static cultivation in the same device or by dynamic cultivation.

摘要

在三维支架上实现细胞黏附对于达到高细胞密度和均匀的细胞分布来说是一项具有挑战性的任务。我们旨在设计一种用于静态接种和培养的三维细胞培养装置(3D-CD),使其能与任何类型的支架配合使用,减少细胞损失并促进营养供应。三维打印技术用于支架和装置的制造。除了测试该装置外,本研究的目的是评估和比较在常氧和低氧条件下,湿支架和干支架的静态和动态接种及培养方法,以及它们对细胞接种效率、细胞分布和细胞增殖等参数的影响。采集人体脂肪组织并在三维打印的聚己内酯支架中培养。进行了微型计算机断层扫描(Micro-CT),并将投影图像重建为横截面图像。我们创建了三维图像以可视化支架内的细胞分布和取向。预湿支架组最有利于细胞附着。三维细胞培养装置(3D-CD)提高了细胞接种效率,几乎没有细胞损失。我们建议,在该装置中进行24小时静态接种,然后在同一装置中进行静态培养或动态培养,可产生最有利的结果。

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