Nydus One Syringe Extruder (NOSE): A Prusa i3 3D printer conversion for bioprinting applications utilizing the FRESH-method
Nydus One Syringe Extruder (NOSE): A Prusa i3 3D printer conversion for bioprinting applications utilizing the FRESH-method
复制标题
DOI：
10.1016/j.ohx.2019.e00069
复制
发表时间：
2019-10-01
期刊：
影响因子：
2.2
通讯作者：
Brand-Saberi, Beate
中科院分区：
文献类型：
--
作者：
Bessler, Nils;Ogiermann, Dennis;Brand-Saberi, Beate
生物打印与其他组织工程技术相结合，是创建工程组织或标准化3D细胞培养模型的一种有前途的方法。它在开发用于药物毒性筛选或个性化医疗方法的无动物模型中具有潜在的应用。在这里，我们报告了全球使用的开源3D打印机（RepRap，Prusa i3）通过用Nydus One Syringe Extruder（NOSE）取代普通塑料挤出机，将其转换为功能性和负担得起的生物打印机。NOSE改进通过采用模块化压模支架概念实现了机械水凝胶挤出以及可调的沉积精度和体积。开发了一种具有成本效益和简单的硬件设计，包括定制软件（称为“合成器”），以证明这种技术可以为更广泛的公众所用。此外，欣顿等人针对同行评审的FRESH方法优化了打印程序，该方法允许打印几何形状复杂的载有细胞的构建体。我们提供了有关新鲜凝胶创建的详细方案，以确保这种最先进方法的重现性。作为概念的证明，HEK293细胞以及小鼠胚胎干细胞（mESC）用于载有细胞的打印。根据细胞来源，存活率从60%到95%不等。使用开源图像分析工具Icy，特别是“斑点检测器”插件进行自动化定量活力分析。详细的方案允许重新创建测定。利用支持浴印刷技术的进一步数据揭示了关于载有细胞的构建体的印刷和停留时间的限制。(C)2019作者爱思唯尔有限公司出版
Bioprinting, combined with other tissue engineering techniques, is a promising method to create engineered tissues or standardized 3D cell culturing models. It has potential applications in the development of animal free models for drug toxicity screenings or for personalized medicine approaches. Here we report the conversion of the worldwide used open source 3D printer (RepRap, Prusa i3) to a functional and affordable bioprinter by substituting the common plastic-extruder with the Nydus One Syringe Extruder (NOSE). The NOSE modification enables mechanical hydrogel extrusion as well as a tunable deposition precision and volume by featuring a modular syringe-holder concept. A cost effective and simple hardware design including a custom software (termed 'composer') was developed to prove that this technique can be made accessible for a broader public. Furthermore, the printing procedure was optimized for the peer-reviewed FRESH-method by Hinton et al. which allows to print geometrical complex cell-laden constructs. We provide a detailed protocol on the creation of the FRESH-gel to ensure the reproducibility of this state of the art method. As a proof of concept HEK293 cells as well as mouse embryonic stem cells (mESC) were utilized for cell-laden printing. Depending on the cellular source the survival rates range from 60% up to 95%. Automatized quantitative viability analysis was performed using the open source image analysis tool Icy, in particular the "spot detector" plugin. A detailed protocol allows the recreation of the assay. Further data utilizing a support bath printing technique reveal limitations regarding the printing and residential time of cell-laden constructs. (C) 2019 The Authors. Published by Elsevier Ltd.