identification for unknown cell sources. It offers potential to conveniently leverage the communication between single cell electroporation for cell electrophysiology study and bulk electroporation
processes used in large scale gene function and therapy practice. In
this contribution, we evaluated the transfection enhancement of
MAE to reporter genes (pMaxGFP and gWizLuc) and their
corresponding siRNA probes. Several adherent and suspension
cell lines were tested to demonstrate its broad effectiveness.
2 Materials
2.1 Plasmid DNA
and siRNA
The DNA plasmids with gWiz luciferase and pMax GFP reporter
genes are purchased from Aldevron, Inc. and Lonza, Inc. respectively. Small interfering RNA (siRNA) used for silencing GFP
(expressed by pMaxGFP) and Luciferase genes are synthesized
by Thermo Scientific (Pittsburgh, PA) and the sequences are as
follows: siRNA for GFP silence, sense strand, 5
0 -CGCAUG
ACCAACAAGAUGAUU-3
0 ; antisense strand, 5
0 -UCAUCUUG
UUGGUCAUGCGGC-3
0 ; Luciferase GL3 Duplex (Luc-siRNA),
sense strand, 5
0 -CUUACGCUGAGUACUUCGA-3
0 ; antisense
strand, 5
0 -UCGAAGUACUCAGCGUAAG-3
0 .
2.2 Cell Culture
NIH 3T3 cells (ATCC, CRL-1658) are maintained in high-glucose
DMEM supplemented with 10% newborn calf serum (NCS), 1%
penicillin and streptomycin, 1% L-glutamine, and 1% sodium pyruvate. K562 cells (ATCC, CCL-243), A549 (ATCC, CCL-185),
HeLa (ATCC, CCL-2), COS-7 (ATCC, CRL-1651), 293T
(ATCC, CRL-3216), and HL-60 (ATCC, CCL-240) are cultured
in RPMI 1640 supplemented with 10% NCS, 100 U/mL penicillin, 100 μg/mL streptomycin, and 100 μg/mL L-glutamine.
Mouse embryonic stem (ATCC, CRL-1934) are cultured on
gelatin-coated tissue culture flasks and maintained in an undifferentiated state using Dulbecco’s Modified Eagle’s Medium (DMEM
with 4.5 g/L D-glucose) supplemented with 15% (v/v) fetal bovine
serum (FBS), 100 U/mL penicillin, 100 μg/mL streptomycin,
0.1 mM nonessential amino acids, 10 ng/mL murine recombinant
leukemia inhibitory factor (LIF), 0.1 mM monothioglycerol,
2 mM L-glutamine (Sigma-Aldrich, St. Louis, MO), and 1 mM
sodium pyruvate (Invitrogen). All cultures are maintained at 37
C
with 5% CO 2 and 100% relative humidity.
2.3 Micropillar Array
Fabrication
Micropillar arrays are fabricated by BioMEMS Technologies.
Briefly, SU-8 photoresist is patterned on a Si (100) wafer via
photolithography. Micropillars (2 μm in diameter with a pitch size
of 2 μm or 6 μm and 4–5 μm in height, Fig. 1b) are defined on a
20-mm  20-mm silicon wafer. Conductive micropillars are made
by further sputter coating the SU-8 pattern with gold. A second
Cell Size-Specific Transfection by Micropillar Array Electroporation
5
processes used in large scale gene function and therapy practice. In
this contribution, we evaluated the transfection enhancement of
MAE to reporter genes (pMaxGFP and gWizLuc) and their
corresponding siRNA probes. Several adherent and suspension
cell lines were tested to demonstrate its broad effectiveness.
2 Materials
2.1 Plasmid DNA
and siRNA
The DNA plasmids with gWiz luciferase and pMax GFP reporter
genes are purchased from Aldevron, Inc. and Lonza, Inc. respectively. Small interfering RNA (siRNA) used for silencing GFP
(expressed by pMaxGFP) and Luciferase genes are synthesized
by Thermo Scientific (Pittsburgh, PA) and the sequences are as
follows: siRNA for GFP silence, sense strand, 5
0 -CGCAUG
ACCAACAAGAUGAUU-3
0 ; antisense strand, 5
0 -UCAUCUUG
UUGGUCAUGCGGC-3
0 ; Luciferase GL3 Duplex (Luc-siRNA),
sense strand, 5
0 -CUUACGCUGAGUACUUCGA-3
0 ; antisense
strand, 5
0 -UCGAAGUACUCAGCGUAAG-3
0 .
2.2 Cell Culture
NIH 3T3 cells (ATCC, CRL-1658) are maintained in high-glucose
DMEM supplemented with 10% newborn calf serum (NCS), 1%
penicillin and streptomycin, 1% L-glutamine, and 1% sodium pyruvate. K562 cells (ATCC, CCL-243), A549 (ATCC, CCL-185),
HeLa (ATCC, CCL-2), COS-7 (ATCC, CRL-1651), 293T
(ATCC, CRL-3216), and HL-60 (ATCC, CCL-240) are cultured
in RPMI 1640 supplemented with 10% NCS, 100 U/mL penicillin, 100 μg/mL streptomycin, and 100 μg/mL L-glutamine.
Mouse embryonic stem (ATCC, CRL-1934) are cultured on
gelatin-coated tissue culture flasks and maintained in an undifferentiated state using Dulbecco’s Modified Eagle’s Medium (DMEM
with 4.5 g/L D-glucose) supplemented with 15% (v/v) fetal bovine
serum (FBS), 100 U/mL penicillin, 100 μg/mL streptomycin,
0.1 mM nonessential amino acids, 10 ng/mL murine recombinant
leukemia inhibitory factor (LIF), 0.1 mM monothioglycerol,
2 mM L-glutamine (Sigma-Aldrich, St. Louis, MO), and 1 mM
sodium pyruvate (Invitrogen). All cultures are maintained at 37
C
with 5% CO 2 and 100% relative humidity.
2.3 Micropillar Array
Fabrication
Micropillar arrays are fabricated by BioMEMS Technologies.
Briefly, SU-8 photoresist is patterned on a Si (100) wafer via
photolithography. Micropillars (2 μm in diameter with a pitch size
of 2 μm or 6 μm and 4–5 μm in height, Fig. 1b) are defined on a
20-mm  20-mm silicon wafer. Conductive micropillars are made
by further sputter coating the SU-8 pattern with gold. A second
Cell Size-Specific Transfection by Micropillar Array Electroporation
5