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Abstract：
Automated microfluidic devices are a promising route towards a point-of-care autologous cell therapy. The initial steps of induced pluripotent stem cell (iPSC) derivation involve transfection and long term cell culture. Integration of these steps would help reduce the cost and footprint of micro-scale devices with applications in cell reprogramming or gene correction. Current examples of transfection integration focus on maximising efficiency rather than viable long-term culture. Here we look for whole process compatibility by integrating automated transfection with a perfused microfluidic device designed for homogeneous culture conditions. The injection process was characterised using fluorescein to establish a LabVIEW-based routine for user-defined automation. Proof-of-concept is demonstrated by chemically transfecting a GFP plasmid into mouse embryonic stem cells (mESCs). Cells transfected in the device showed an improvement in efficiency (34%, n = 3) compared with standard protocols (17.2%, n = 3). This represents a first step towards microfluidic processing systems for cell reprogramming or gene therapy.
摘要：
自动化微流体装置是实现定点护理自体细胞治疗的有希望的途径。诱导多能干细胞(iPSC)衍生的初始步骤涉及转染和长期细胞培养。这些步骤的整合将有助于降低微尺度设备在细胞重编程或基因校正中的应用的成本和占地面积。转染整合的当前实例集中于最大化效率而不是可行的长期培养。在这里，我们通过将自动转染与设计用于均质培养条件的灌注微流体装置集成来寻找整个过程的兼容性。使用荧光素对注射过程进行表征，以建立基于LabVIEW的例程，以实现用户定义的自动化。通过将GFP质粒化学转染到小鼠胚胎干细胞（mESC）中来证明概念证明。在设备中转染的细胞显示效率提高（34％，n=3）与标准协议（17.2%，n=3)。这代表了朝向用于细胞重编程或基因治疗的微流体处理系统的第一步。
