Design and validation of an electroporation-capable random positioning machine for the study of cell membrane permeabilization in simulated microgravity

Biomanufacturing is being explored as a potential solution for improved space logistics in austere environments. One potentially useful tool is electroporation-the delivery of exogenous DNA into cells using pulsed electric fields to increase cell membrane permeability. Electroporation has the potential to enable tunable bioreactors in a space environment through the targeted delivery of nucleic acids encoding proteins of interest and may also present a solution for disinfection, purification, and biomolecule extraction without requiring disposable reagents. Unfortunately, electroporation has not been studied using cells adapted to microgravity conditions, and it is possible that well-known cellular adaptations to microgravity will alter electroporation outcomes. Studying this phenomenon is complicated by the fact that electroporating cells in common microgravity simulators, such as the rotating wall vessel and the random positioning machine (RPM), is currently not feasible. In this work, we design and validate a custom, electroporation-capable RPM. The device consists of two rotating frames, which continuously reorient a biological sample relative to the direction of gravity, while also allowing for the delivery of intense electric pulses required for electroporation. The confirmation of the RPM's simulation of microgravity was assessed using an accelerometer. Once validated, a proof-of-concept experiment was performed on human T-lymphocytes to assess the effect of microgravity adaptation on membrane permeability following electroporation. Cell permeability after electroporation decreased after simulated microgravity exposure, suggesting that cellular adaptations to microgravity may alter electroporation outcomes.