Transfection in perfused microfluidic cell culture devices: A case study

[Display omitted] •Automated chemical transfection in a microfluidic device for long-term homogeneous cell culture.•Transfection reagent mixture uniformly exposed to cells growing in the device.•Embryonic stem cells transfected with GFP show improved efficiency and an increased median fluorescence intensity on-chip relative to 24 well plate. 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.