Micro-/nanofluidics based cell electroporation.

Non-viral gene delivery has been extensively explored as the replacement for viral systems. Among various non-viral approaches, electroporation has gained increasing attention because of its easy operation and no restrictions on probe or cell type. Several effective systems are now available on the market with reasonably good gene delivery performance. To facilitate broader biological and medical applications, micro-/nanofluidics based technologies were introduced in cell electroporation during the past two decades and their advances are summarized in this perspective. Compared to the commercially available bulk electroporation systems, they offer several advantages, namely, (1) sufficiently high pulse strength generated by a very low potential difference, (2) conveniently concentrating, trapping, and regulating the position and concentration of cells and probes, (3) real-time monitoring the intracellular trafficking at single cell level, and (4) flexibility on cells to be transfected (from single cell to large scale cell population). Some of the micro-devices focus on cell lysis or fusion as well as the analysis of cellular properties or intracellular contents, while others are designed for gene transfection. The uptake of small molecules (e.g., dyes), DNA plasmids, interfering RNAs, and nanoparticles has been broadly examined on different types of mammalian cells, yeast, and bacteria. A great deal of progress has been made with a variety of new micro-/nanofluidic designs to address challenges such as electrochemical reactions including water electrolysis, gas bubble formation, waste of expensive reagents, poor cell viability, low transfection efficacy, higher throughput, and control of transfection dosage and uniformity. Future research needs required to advance micro-/nanofluidics based cell electroporation for broad life science and medical applications are discussed.

[1]  O Orwar,et al.  Electroporation of single cells and tissues with an electrolyte-filled capillary. , 2001, Analytical chemistry.

[2]  Brian E. Henslee,et al.  Electroporation dependence on cell size: optical tweezers study. , 2011, Analytical chemistry.

[3]  M. Rols,et al.  Direct visualization at the single-cell level of electrically mediated gene delivery , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Y. Zhan,et al.  Vortex-assisted DNA delivery. , 2010, Lab on a chip.

[5]  H. C. Mastwijk,et al.  Electroporation of cells in microfluidic devices: a review , 2006, Analytical and bioanalytical chemistry.

[6]  Brian E. Henslee,et al.  Gene transfection of mammalian cells using membrane sandwich electroporation. , 2007, Analytical chemistry.

[7]  Limu Wang,et al.  Electroporation of micro‐droplet encapsulated HeLa cells in oil phase , 2010, Electrophoresis.

[8]  Effect of geometry on impedance of cell suspended media in electrically mediated molecule uptake using a microstructure , 2008 .

[9]  P. Cullen,et al.  Electroporation can cause artefacts due to solubilization of cations from the electrode plates. Aluminum ions enhance conversion of inositol 1,3,4,5-tetrakisphosphate into inositol 1,4,5-trisphosphate in electroporated L1210 cells. , 1991, The Biochemical journal.

[10]  I. Schmidt-Wolf,et al.  Regulatable systemic production of monoclonal antibodies by in vivo muscle electroporation , 2004, Genetic vaccines and therapy.

[11]  Mario R. Capecchi,et al.  High efficiency transformation by direct microinjection of DNA into cultured mammalian cells , 1980, Cell.

[12]  S. Yamanaka,et al.  Induction of pluripotent stem cells from fibroblast cultures , 2007, Nature Protocols.

[13]  Tapobrata Panda,et al.  Electroporation: basic principles, practical considerations and applications in molecular biology , 1997 .

[14]  Chang Lu,et al.  Electroporation of mammalian cells in a microfluidic channel with geometric variation. , 2006, Analytical chemistry.

[15]  Yu-Cheng Lin,et al.  Electroporation microchips for continuous gene transfection , 2001 .

[16]  N. Munce,et al.  Microfabricated system for parallel single-cell capillary electrophoresis. , 2004, Analytical chemistry.

[17]  Zhao-Lun Fang,et al.  Integration of single cell injection, cell lysis, separation and detection of intracellular constituents on a microfluidic chip. , 2004, Lab on a chip.

[18]  J Teissié,et al.  An experimental evaluation of the critical potential difference inducing cell membrane electropermeabilization. , 1993, Biophysical journal.

[19]  M. McClain,et al.  Microfluidic devices for the high-throughput chemical analysis of cells. , 2003, Analytical chemistry.

[20]  Y. Tai,et al.  A micro cell lysis device , 1999 .

[21]  J Teissié,et al.  Direct observation in the millisecond time range of fluorescent molecule asymmetrical interaction with the electropermeabilized cell membrane. , 1997, Biophysical journal.

[22]  S. Yamanaka,et al.  Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors , 2006, Cell.

[23]  Michelle Khine,et al.  Electrophoresis-assisted single-cell electroporation for efficient intracellular delivery , 2008, Biomedical microdevices.

[24]  Tilak Jain,et al.  Bio-chip for spatially controlled transfection of nucleic acid payloads into cells in a culture. , 2007, Lab on a chip.

[25]  Kazuhiro Sudo,et al.  Efficient Transfection of Embryonic and Adult Stem Cells , 2004, Stem cells.

[26]  Astrid Hamm,et al.  Efficient transfection method for primary cells. , 2002, Tissue engineering.

[27]  Weixiong Wang,et al.  Semicontinuous flow electroporation chip for high-throughput transfection on mammalian cells. , 2009, Analytical chemistry.

[28]  R. Lee,et al.  Surfactant-induced sealing of electropermeabilized skeletal muscle membranes in vivo. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[29]  R Pepperkok,et al.  The many ways to cross the plasma membrane , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[30]  Luke P. Lee,et al.  Single-cell electroporation arrays with real-time monitoring and feedback control. , 2007, Lab on a chip.

[31]  E. Neumann,et al.  Gene transfer into mouse lyoma cells by electroporation in high electric fields. , 1982, The EMBO journal.

[32]  Hao Lin,et al.  Low-frequency ac electroporation shows strong frequency dependence and yields comparable transfection results to dc electroporation. , 2012, Journal of controlled release : official journal of the Controlled Release Society.

[33]  Tobias Meyer,et al.  Suspended-drop electroporation for high-throughput delivery of biomolecules into cells , 2008, Nature Methods.

[34]  Jing Fang,et al.  Electroporation based on hydrodynamic focusing of microfluidics with low dc voltage , 2010, Biomedical microdevices.

[35]  Hidehiro Oana,et al.  Electroporation through a micro-fabricated orifice and its application to the measurement of cell response to external stimuli , 2006 .

[36]  Owe Orwar,et al.  Scanning electroporation of selected areas of adherent cell cultures. , 2007, Analytical chemistry.

[37]  J. Itskovitz‐Eldor,et al.  Nucleofection of human embryonic stem cells. , 2005, Stem cells and development.

[38]  Shengnian Wang,et al.  Targeted nanoparticles enhanced flow electroporation of antisense oligonucleotides in leukemia cells. , 2010, Biosensors & bioelectronics.

[39]  Claire Dalmay,et al.  Design and realization of a microfluidic device devoted to the application of ultra-short pulses of electrical field to living cells , 2011 .

[40]  J. Gehl,et al.  Electroporation: theory and methods, perspectives for drug delivery, gene therapy and research. , 2003, Acta physiologica Scandinavica.

[41]  Shengnian Wang,et al.  Micronozzle array enhanced sandwich electroporation of embryonic stem cells. , 2010, Analytical chemistry.

[42]  Dong-Chul Han,et al.  Electrotransfection of mammalian cells using microchannel-type electroporation chip. , 2004, Analytical chemistry.

[43]  S Y Ho,et al.  Electroporation of cell membranes: a review. , 1996, Critical reviews in biotechnology.

[44]  Yu-Cheng Lin,et al.  Simulation and experimental demonstration of the electric field assisted electroporation microchip for in vitro gene delivery enhancement. , 2004, Lab on a chip.

[45]  Boris Rubinsky,et al.  Instantaneous, quantitative single-cell viability assessment by electrical evaluation of cell membrane integrity with microfabricated devices , 2003 .

[46]  Yong Huang,et al.  Microfabricated electroporation chip for single cell membrane permeabilization , 2001 .

[47]  D. Chang,et al.  High-efficiency gene transfection by in situ electroporation of cultured cells. , 1991, Biochimica et biophysica acta.

[48]  O Orwar,et al.  Altering the biochemical state of individual cultured cells and organelles with ultramicroelectrodes. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[49]  Boris Rubinsky,et al.  Micro-Electroporation: Improving the Efficiency and Understanding of Electrical Permeabilization of Cells , 1999 .

[50]  A. Keating,et al.  Stable expression of selectable genes introduced into human hematopoietic stem cells by electric field-mediated DNA transfer. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[51]  Luke P. Lee,et al.  A single cell electroporation chip. , 2005, Lab on a chip.

[52]  G. Timp,et al.  Using a nanopore for single molecule detection and single cell transfection. , 2012, The Analyst.

[53]  Bo Yu,et al.  Nanochannel electroporation delivers precise amounts of biomolecules into living cells. , 2011, Nature nanotechnology.

[54]  Yu Sun,et al.  Microfluidic approaches for gene delivery and gene therapy. , 2011, Lab on a chip.

[55]  Yong Huang,et al.  Flow-through micro-electroporation chip for high efficiency single-cell genetic manipulation , 2003 .

[56]  James A Thomson,et al.  Neural differentiation of human induced pluripotent stem cells follows developmental principles but with variable potency , 2010, Proceedings of the National Academy of Sciences.

[57]  Daniel W. Pack,et al.  Design and development of polymers for gene delivery , 2005, Nature Reviews Drug Discovery.

[58]  Y. Zhan,et al.  Electroporation of cells in microfluidic droplets. , 2009, Analytical chemistry.

[59]  E. Yeung Study of single cells by using capillary electrophoresis and native fluorescence detection. , 1999, Journal of chromatography. A.

[60]  T. Tsong,et al.  Reversible and irreversible modification of erythrocyte membrane permeability by electric field. , 1985, Biochimica et biophysica acta.

[61]  Regina Luttge,et al.  Apoptotic cell death dynamics of HL60 cells studied using a microfluidic cell trap device. , 2005, Lab on a chip.

[62]  M. Rols,et al.  Temperature effects on electrotransfection of mammalian cells. , 1994, Nucleic acids research.

[63]  A. van den Berg,et al.  Gene transfer and protein dynamics in stem cells using single cell electroporation in a microfluidic device. , 2008, Lab on a chip.

[64]  Chikashi Nakamura,et al.  A molecular delivery system by using AFM and nanoneedle. , 2005, Biosensors & bioelectronics.

[65]  Chikashi Nakamura,et al.  Nanoscale operation of a living cell using an atomic force microscope with a nanoneedle. , 2005, Nano letters.

[66]  Chang Lu,et al.  Microfluidic cell fusion under continuous direct current voltage , 2006 .

[67]  Dharmakeerthi Nawarathna,et al.  Localized electroporation and molecular delivery into single living cells by atomic force microscopy , 2008 .

[68]  Chang Lu,et al.  A microfluidic flow-through device for high throughput electrical lysis of bacterial cells based on continuous dc voltage. , 2006, Biosensors & bioelectronics.

[69]  Keunchang Cho,et al.  A multi-channel electroporation microchip for gene transfection in mammalian cells. , 2007, Biosensors & bioelectronics.

[70]  Chikashi Nakamura,et al.  Gene expression using an ultrathin needle enabling accurate displacement and low invasiveness. , 2005, Biochemical and biophysical research communications.

[71]  Chang Lu,et al.  Microfluidic electroporation for delivery of small molecules and genes into cells using a common DC power supply , 2008, Biotechnology and bioengineering.

[72]  Chang Lu,et al.  High‐throughput and real‐time study of single cell electroporation using microfluidics: Effects of medium osmolarity , 2006, Biotechnology and bioengineering.

[73]  C. Bertozzi,et al.  A cell nanoinjector based on carbon nanotubes , 2007, Proceedings of the National Academy of Sciences.

[74]  G. A. Hofmann,et al.  Medical applications of electroporation , 2000 .

[75]  Wei Wang,et al.  A laminar flow electroporation system for efficient DNA and siRNA delivery. , 2011, Analytical chemistry.

[76]  Jingjiao Guan,et al.  Large laterally ordered nanochannel arrays from DNA combing and imprinting. , 2010, Advanced materials.

[77]  Ali Khademhosseini,et al.  Microscale electroporation: challenges and perspectives for clinical applications. , 2009, Integrative biology : quantitative biosciences from nano to macro.

[78]  Tadej Kotnik,et al.  Sensitivity of transmembrane voltage induced by applied electric fields—A theoretical analysis , 1997 .

[79]  K. Jensen,et al.  A microfluidic electroporation device for cell lysis. , 2005, Lab on a chip.

[80]  R. Morgenstern,et al.  Efficient gene transfer into murine embryonic stem cells by nucleofection , 2004, Biotechnology Letters.

[81]  Keunchang Cho,et al.  A novel electroporation method using a capillary and wire-type electrode. , 2008, Biosensors & bioelectronics.