reprogramming of an adult somatic cell to induced pluripotent
stem cells (iPSCs) [6]. Unlike embryonic stem cells (ESCs),
iPSCs have the ability for consistent gene expression even after
extended passaging as well as can form all somatic lineages
[7]. On the other hand, human ESCs exhibit low efficiencies in
gene targeting and heterogeneity in differentiation potential
among different human ESC lines [7]. Thus, cell reprogramming
of somatic cell to iPSCs using OSKM bypasses the need to destruct
the embryo as well as the related ethical concerns. Since iPSCs are
reprogrammed cells from the same patient, they eliminate immune
system incompatibility issues. In addition, iPSCs are ideal to model
human disease in vitro [8] and an effective therapeutic tool in
regenerative medicine. However, the widely used viral transduction
for delivering OSKM factors instigates mutagenesis, which compromises the use of iPSCs for human clinical trials [9].
In the third protocol, the efficacy of the 3D NEP was tested for
its application in transfecting some special cells which originally
were difficult to be transfected. Transfection of cardiomyocytes
has always remained a challenge since the available electroporation
methods activate the ion channels, leading to the abnormal influx
of ions. Therefore, cardiomyocytes were selected as the cells to be
transfected in order to emphasize the significance of the NEP
device. The significance of the device for dosage controllability
was manifested by injecting miR29 into primary cardiomyocytes.
Loss of miR29 via a Smad3-dependent mechanism is associated
with cardiac fibrosis in a hypertensive heart [10]. When miR29 is
downregulated, it induces the expression of collagen, whereas its
overexpression reduces collagen expression [11]. Therefore,
miR29, if replenished at a correct dose, can be a potential target
to treat tissue fibrosis.
In summary, this novel 3D NEP system was utilized to
deliver macromolecules into living cells without compromising
the transfection efficiency and cell viability. Adoptive immunotherapy, cell reprogramming and miR-induced gene therapy were
successfully demonstrated by injecting CAR plasmid into NK
cells, OSKM plasmids into mouse embryonic fibroblast and
miR29 into cardiomyocytes respectively. Finally, the data from
the transfected cells was analyzed to measure the transfection
efficiency, dosage controllability, as well as tested the performance of the device. Overall, this high-throughput 3D NEP
system provides an innovative and medically valuable platform
with uniform and reliable cellular transfection and cell
reprogramming [12].
3D Nanochannel Electroporation for Macromolecular Nucleotide Delivery
71
stem cells (iPSCs) [6]. Unlike embryonic stem cells (ESCs),
iPSCs have the ability for consistent gene expression even after
extended passaging as well as can form all somatic lineages
[7]. On the other hand, human ESCs exhibit low efficiencies in
gene targeting and heterogeneity in differentiation potential
among different human ESC lines [7]. Thus, cell reprogramming
of somatic cell to iPSCs using OSKM bypasses the need to destruct
the embryo as well as the related ethical concerns. Since iPSCs are
reprogrammed cells from the same patient, they eliminate immune
system incompatibility issues. In addition, iPSCs are ideal to model
human disease in vitro [8] and an effective therapeutic tool in
regenerative medicine. However, the widely used viral transduction
for delivering OSKM factors instigates mutagenesis, which compromises the use of iPSCs for human clinical trials [9].
In the third protocol, the efficacy of the 3D NEP was tested for
its application in transfecting some special cells which originally
were difficult to be transfected. Transfection of cardiomyocytes
has always remained a challenge since the available electroporation
methods activate the ion channels, leading to the abnormal influx
of ions. Therefore, cardiomyocytes were selected as the cells to be
transfected in order to emphasize the significance of the NEP
device. The significance of the device for dosage controllability
was manifested by injecting miR29 into primary cardiomyocytes.
Loss of miR29 via a Smad3-dependent mechanism is associated
with cardiac fibrosis in a hypertensive heart [10]. When miR29 is
downregulated, it induces the expression of collagen, whereas its
overexpression reduces collagen expression [11]. Therefore,
miR29, if replenished at a correct dose, can be a potential target
to treat tissue fibrosis.
In summary, this novel 3D NEP system was utilized to
deliver macromolecules into living cells without compromising
the transfection efficiency and cell viability. Adoptive immunotherapy, cell reprogramming and miR-induced gene therapy were
successfully demonstrated by injecting CAR plasmid into NK
cells, OSKM plasmids into mouse embryonic fibroblast and
miR29 into cardiomyocytes respectively. Finally, the data from
the transfected cells was analyzed to measure the transfection
efficiency, dosage controllability, as well as tested the performance of the device. Overall, this high-throughput 3D NEP
system provides an innovative and medically valuable platform
with uniform and reliable cellular transfection and cell
reprogramming [12].
3D Nanochannel Electroporation for Macromolecular Nucleotide Delivery
71