Introduction
Multiple myeloma is a differentiated plasma cell malignancy and is the
most common hematological malignancy after non-Hodgkin’s Lymphoma
(Siegel, Miller, & Jemal, 2016). This disorder accounts for
1% of all malignancies and 10% of blood malignancies. The disease is
characterized by the clonal proliferation of malignant plasma cells in
the bone marrow, which results in the overproduction of monoclonal
immunoglobulins in the blood and urine of patients. In addition,
patients develop significant bone lesions that endanger patients’
quality and survival time (Kyle et al., 2003)(Palumbo & Anderson,
2011). Multiple myeloma has been an obstructive disease to treat. The
median overall survival was approximately 3 years until the late 1990s.
The introduction of high-dose therapy, particularly, and autologous stem
cell transplantation, as well as the development of new drugs, such as
immunomodulatory drugs (IMiDs) and proteasome inhibitors lead to the
improvement in survival. However, finally most MM patients relapse which
reveal that there is a need for novel treatment approaches (Franssen,
Mutis, Lokhorst, & van de Donk, 2019).
Ex vivo manipulation of cells is crucial for the success of cell-based
therapies, such as chimeric antigen receptor T cells for the treatment
of cancers but challenges in delivery technologies have limited this
approach (Cox, Platt, & Zhang, 2015).Current method for manufacturing
gene-modified cells requires the use of viruses which is time consuming
and expensive (Wang & Rivière, 2016). In addition, viral gene delivery
system needs safety and monitoring of patients after treatment (Levine,
Miskin, Wonnacott, & Keir, 2017).non-viral gene delivery is an
alternative to viral- based gene modified cell therapy(GMCT) because of
low cost, easy to scale and brief lead time. Nucleofection is one of the
routine non-viral techniques that can transfer genetic materials to
broad range of cells. This method is easy, rapid, efficient that
resulted in greater than 80% expression of transferred constructs in
human T cells (Tchou et al., 2017). However, because of reduced cell
viability and disruptions on global gene expression, cytokine
production, lineage markers, and in vivo function of cells, now is less
than ideal for GMCT (Escoffre et al., 2009)´(Rosazza, Haberl Meglic,
Zumbusch, Rols, & Miklavcic, 2016). Thus, novel non-viral intracellular
delivery strategies are required to overcome limitations of conventional
methods in efficient and nontoxic transfer of various macromolecules to
the immune cell engineering.
Microfluidic technology is the science of using low amounts of liquid
flow (microliter, nanoliter) in the synthesized microsystems. The
applications of this approach in biology and medicine have led to the
creation of a new research direction. Since the beginning of this
technology, in the 1990’s, almost concomitant with the advent of
micro-scale chemistry techniques, microfluidics have grown rapidly and
persisted with the promise of evolving conventional laboratory analysis
techniques (Loessner et al., 2010).Benefits of this technique include
significant reduction in material and sample consumption, increased
speed and sensitivity to reactions, separation and detection, reduced
cost and time of operation, and importantly in the three-dimensional
imitation of cell environment (Whitesides, 2006).Since gene transfer
with conventional systems requires complex functional processes and
lacks the sensitivity and specificity as well as low viability and
efficiency, rapid advancement in the field of microfluidic knowledge
opens new window for gene transfer and Gene therapy .In general, the
microfluidic method has facilitated the transfer of macromolecules into
the cell by scaling the channel and cell dimensions(Xia et al., 2010).
Since intracellular transfer of biomolecules such as DNA and siRNA to
immune cells is difficult, so here, we describe the development and use
of our user-friendly microfluidic transfection device for hydro
dynamically intracellular delivery of DNA into myeloma cells using
serpentine channels. we delivered about 11.0 Kb DNA construct expressing
green fluorescent protein (GFP) and anti-BCMA marker into human Myeloma
cell and showed high levels of cell viability, and gene expression. This
work provides comparative functional consequences of nucleofection as a
2D gene transfer strategy relative to an innovative mechanical delivery
strategy as 3D gene delivery approach.