Klaus Kreuels, Carina Schemmer, Arnold Gillner, Maximilian Frederick Flesch, Arnold Gillner
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引用次数: 0
摘要
本研究提出了一种三维(3D)细胞培养的新方法,该方法使用由3D打印支架结构和灌注通道组成的双组分系统。采用热紫外立体光刻(SLA)系统进行3d打印,制备了一种总尺寸为9mm x 9mm x 1mm的聚合物支架结构,该支架由立方晶格组成,腹板厚度为200 μm,孔径为600 μm。采用相同的体系和材料,3d打印出内径为800 μm、壁孔为300 μm的灌注通道供细胞培养基供应。用数码显微镜对支架的打印精度进行了研究。采用mtt法和活/死染色评估材料的细胞毒性。随后,将3T3-成纤维细胞植入基于纤维蛋白的水凝胶中,然后使用静水压力驱动流系统进行静态或被动灌注调节。结果表明,该支架结构和灌注通道的制备精度高,稳定性好,可使支架内细胞通过灌注通道获得营养和氧气供应。这种方法有潜力在组织工程和再生医学应用的更大结构中提供营养。
3D-printed scaffolds with perfusable channels for low-cost large construct 3D cell culture
Abstract This study presents a novel approach for threedimensional (3D) cell culture using a two-component system consisting of a 3D-printed scaffold structure combined with a perfusion channel. A polymeric scaffold structure with an overall size of 9 mm x 9 mm x 1 mm composed of a cubic lattice with a web thickness of 200 μm and pore size of 600 μm was 3D-printed using a hot UV-stereolithography (SLA) system. The perfusion channel with an inner diameter of 800 μm and channel wall pores of 300 μm for cell culture medium supply was 3D-printed with the same system and material. Scaffolds were investigated with respect to the printing accuracy by digital microscopy. Cytotoxicity of the materials was assessed using MTT-assay and Live/Dead staining. Scaffold were subsequently seeded with 3T3- fibroblasts within a fibrin-based hydrogel and then conditioned either statically or under passive perfusion using a hydrostatic pressure driven flow system. The results show that the scaffold structure and perfusion channel can be produced with high accuracy and stability allowing a supply of nutrient and oxygen via perfusion channel to the cells within the scaffold. This approach has potential for nutrient supply within larger constructs for tissue engineering and regenerative medicine applications.