Project R-13310

In-flow cell electroporation with single-cell level control using impedance spectroscopy (Research)

Induced Pluripotent Stem Cells (iPSC) have been lauded as a breakthrough in regenerative medicine. The collection and reprogramming of patient cells enable the in vitro creation of patient-specific tissues, promising future treatments for e.g. Parkinson's disease, spinal cord injury and macular degeneration. However, current technologies suffer from low reprogramming efficiencies, preventing their adoption in the clinic. A key need is the robust and stoichiometric delivery of multiple transcription factors into the cells. Reliable delivery of controlled doses of large plasmids would be a crucial breakthrough. Indeed, previous studies have shown 1000-fold improvement in yield for single-cell transfection compared to bulk electroporation and faster expression of biomarkers than upon bulk electroporation and viral transfection. The focus of this PhD is to develop a microfluidic device that will electroporate cells with single-cell level control. In this microfluidic chip, the cells experience a well-controlled field strength while they pass electrodes, thus achieving highly uniform pore formation in the cell membrane. Electroporation is combined with impedance measurements on chip to optimize the electroporation settings for cell viability and genetic modification. It will also be investigated if measurements of cell properties such as cell size with impedance sensing before electroporation can further optimize electroporation performance. The student will start his/her research by establishing an electro-fluidic model for single-cell electroporation and create device designs according to the model. By fabricating simple proof-of-concept devices and characterizing them, the student will validate the model and select optimized designs for final working devices, which will be fabricated in imec FAB. After device fabrication, the student will characterize the devices and validate transfection of cells. This topic will be co-supervised and supported by a team of physicists, biochemists, and engineers in University Hasselt and the imec life sciences department.

16 September 2022 - 15 September 2024