Raimes William, Rubi Mathieu, Super Alexandre, Marques Marco P C, Veraitch Farlan, Szita Nicolas
Department of Biochemical Engineering, University College London, Bernard Katz Building, Gordon Street, London WC1H 0AH, United Kingdom.
Process Biochem. 2017 Aug;59(Pt B):297-302. doi: 10.1016/j.procbio.2016.09.006.
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.
自动化微流控设备是实现即时自体细胞治疗的一条很有前景的途径。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。整合这些步骤将有助于降低微尺度设备在细胞重编程或基因校正应用中的成本和占地面积。当前转染整合的例子侧重于最大化效率而非可行的长期培养。在这里,我们通过将自动转染与为均匀培养条件设计的灌注微流控设备相结合来寻求整个过程的兼容性。使用荧光素对注射过程进行表征,以建立基于LabVIEW的用户定义自动化程序。通过将绿色荧光蛋白(GFP)质粒化学转染到小鼠胚胎干细胞(mESC)中来证明概念验证。与标准方案(17.2%,n = 3)相比,在该设备中转染的细胞效率有所提高(34%,n = 3)。这代表了朝着用于细胞重编程或基因治疗的微流控处理系统迈出的第一步。