Chapter 1
Cell Size-Specific Transfection by Micropillar Array
Electroporation
Xuan Liu, Yingbo Zu, and Shengnian Wang
Abstract
In this study, we reported a new micropillar array electroporation (MAE) platform to accomplish largescale, size-specific DNA and RNA delivery to mammalian cells for nanomedicine. By introducing wellpatterned micropillar array on the electrode surface, the number of micropillars each cell faces varies with
the surface area of cell membrane or the size of cells. In this way, cell size-specific electroporation is
conveniently done on a large population of cells in despite of their random locations between the two
electrodes. The enhancement of this MAE system on the delivery of DNA and RNA probes without
sacrifice of cell viability is demonstrated with an average increase of 2.5 to 3-fold on the transfection
efficiency of DNA plasmids and additional knockdown of the targeted protein 10–55% more in siRNA
delivery when compared to that using a commercial electroporation system. This MAE system works like
many single cell electroporation are carried out in parallel, showing potential to bridge the gap between
single cell electrophysiology study and in vitro electroporation to a large population of cells.
Key words Electroporation, Gene delivery, Transfection enhancement, Micropillar array, Microstructured electrode
1 Introduction
Among various nonviral delivery approaches, electroporation is a
simple and quick delivery tool to make the subjected cell membrane
transiently permeable for quickly uptake of exogenous probes
[1]. Its performance relies on not only the amplitude, duration,
and number of high-voltage pulses that are applied to the cell
membrane but also cell size, the permeability of cell membrane,
and its orientation to the electric field [2]. A number of microscale/
nanoscale electroporation systems have been explored in the past
decade to tackle the high-voltage issues (e.g., water hydrolysis,
leading to pH changes and bubble burst damage [3, 4]) through
closely patterned electrode pairs and/or sophisticated focusing of
the applied electric pulses [5–7]. These microdevices open new
routes towards the elimination of many electroporation induced
Shulin Li et al. (eds.), Electroporation Protocols: Microorganism, Mammalian System, and Nanodevice,
Methods in Molecular Biology, vol. 2050, https://doi.org/10.1007/978-1-4939-9740-4_1,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
3
Cell Size-Specific Transfection by Micropillar Array
Electroporation
Xuan Liu, Yingbo Zu, and Shengnian Wang
Abstract
In this study, we reported a new micropillar array electroporation (MAE) platform to accomplish largescale, size-specific DNA and RNA delivery to mammalian cells for nanomedicine. By introducing wellpatterned micropillar array on the electrode surface, the number of micropillars each cell faces varies with
the surface area of cell membrane or the size of cells. In this way, cell size-specific electroporation is
conveniently done on a large population of cells in despite of their random locations between the two
electrodes. The enhancement of this MAE system on the delivery of DNA and RNA probes without
sacrifice of cell viability is demonstrated with an average increase of 2.5 to 3-fold on the transfection
efficiency of DNA plasmids and additional knockdown of the targeted protein 10–55% more in siRNA
delivery when compared to that using a commercial electroporation system. This MAE system works like
many single cell electroporation are carried out in parallel, showing potential to bridge the gap between
single cell electrophysiology study and in vitro electroporation to a large population of cells.
Key words Electroporation, Gene delivery, Transfection enhancement, Micropillar array, Microstructured electrode
1 Introduction
Among various nonviral delivery approaches, electroporation is a
simple and quick delivery tool to make the subjected cell membrane
transiently permeable for quickly uptake of exogenous probes
[1]. Its performance relies on not only the amplitude, duration,
and number of high-voltage pulses that are applied to the cell
membrane but also cell size, the permeability of cell membrane,
and its orientation to the electric field [2]. A number of microscale/
nanoscale electroporation systems have been explored in the past
decade to tackle the high-voltage issues (e.g., water hydrolysis,
leading to pH changes and bubble burst damage [3, 4]) through
closely patterned electrode pairs and/or sophisticated focusing of
the applied electric pulses [5–7]. These microdevices open new
routes towards the elimination of many electroporation induced
Shulin Li et al. (eds.), Electroporation Protocols: Microorganism, Mammalian System, and Nanodevice,
Methods in Molecular Biology, vol. 2050, https://doi.org/10.1007/978-1-4939-9740-4_1,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
3