Mechanoporation: Toward Single Cell Approaches

Kumar, Amogh; Mohan, L.; Shinde, Pallavi; Chang, Hwan‐You; Nagai, Masao; Santra, Tuhin Subhra

doi:10.1007/978-981-10-4857-9_3-1

Cited by 9 publications

(9 citation statements)

References 65 publications

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“…Irreversible electroporation has proven to be an issue for electroporation as perpetual permeabilization of the cell membrane caused by electrical pulse can lead to substantial cell death and compromises to cell viability 18 . The usage of electric fields to enable transfection causes high toxicity and low cell viability; with cell viability compromised, it is difficult to scale-up the application via this mode of transfection 19 – 21 . Chemical-based transfections, the typical alternative to viral vectors and electroporation, have come up short in transfecting primary cells, including natural killer (NK) and T cells.…”

Section: Introductionmentioning

confidence: 99%

“…Mechanoporation, the delivery of cargo through transient pores created on the cell membrane by mechanical forces, offers a new avenue that can overcome the weaknesses of current transfection methods. Examples of such mechanical forces are shear stress, puncturing or compressions applied to the cell membrane, which can be generated by manipulating cells through microfluidic circuits or onto microneedles 19 . A first example of a mechanoporation method employed in gene therapy is microinjection, a technique where payload is precisely delivered into cells with high accuracy with micropipettes or nanoneedles.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Microfluidic transfection of mRNA into human primary lymphocytes and hematopoietic stem and progenitor cells using ultra-fast physical deformations

Loo¹,

Sicher²,

Goff³

et al. 2021

Sci Rep

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Messenger RNA (mRNA) delivery provides gene therapy with the potential to achieve transient therapeutic efficacy without risk of insertional mutagenesis. Amongst other applications, mRNA can be employed as a platform to deliver gene editing molecules, to achieve protein expression as an alternative to enzyme replacement therapies, and to express chimeric antigen receptors (CARs) on immune cells for the treatment of cancer. We designed a novel microfluidic device that allows for efficient mRNA delivery via volume exchange for convective transfection (VECT). In the device, cells flow through a ridged channel that enforces a series of ultra-fast and large intensity deformations able to transiently open pores and induce convective transport of mRNA into the cell. Here, we describe efficient delivery of mRNA into T cells, natural killer (NK) cells and hematopoietic stem and progenitor cells (HSPCs), three human primary cell types widely used for ex vivo gene therapy applications. Results demonstrate that the device can operate at a wide range of cell and payload concentrations and that ultra-fast compressions do not have a negative impact on T cell function, making this a novel and competitive platform for the development of ex vivo mRNA-based gene therapies and other cell products engineered with mRNA.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Microfluidic transfection of mRNA into human primary lymphocytes and hematopoietic stem and progenitor cells using ultra-fast physical deformations

Loo¹,

Sicher²,

Goff³

et al. 2021

Sci Rep

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“…Furthermore, single cell separation and diagnosis techniques are discussed using microfluidic Bio-MEMS devices integrated with photomechanical, laser capture micro dissection, and nano-capillary chip technologies. Single cell therapeutic and analysis techniques are performed using electroporation [ 32 ], optoporation, mechanoporation [ 33 ], capillary electrophoresis coupled with laser-induced fluorescence detection, flow cytometry, mass spectrometry, high-resolution microscopy, the resonating microelectromechanical system (MEMS), electro neural nanowire device, and nanostraw system. Thus, this review article intends to provide an overview of currently available techniques for single cell manipulation, analysis, and their applications in diagnostic as well as therapeutic development.…”

Section: Introductionmentioning

confidence: 99%

Current Trends of Microfluidic Single-Cell Technologies

Shinde

Mohan

Kumar

et al. 2018

IJMS

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The investigation of human disease mechanisms is difficult due to the heterogeneity in gene expression and the physiological state of cells in a given population. In comparison to bulk cell measurements, single-cell measurement technologies can provide a better understanding of the interactions among molecules, organelles, cells, and the microenvironment, which can aid in the development of therapeutics and diagnostic tools. In recent years, single-cell technologies have become increasingly robust and accessible, although limitations exist. In this review, we describe the recent advances in single-cell technologies and their applications in single-cell manipulation, diagnosis, and therapeutics development.

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“…They can manipulate and control fluids in the range of micro to pico-liters, thus reducing sample loss, providing susceptible analysis in the miniaturized microfluidic systems [ 14 , 15 , 17 ]. The micro/nanofluidic devices are not only designed and fabricated to fulfill the needs of various single-cell manipulation, separation, trapping isolation and lysis, but also used for electrical, mechanical, optical, biochemical characterization, as well as for therapeutic and diagnostic purposes [ 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 ]. Figure 1 shows worldwide microfluidic single-cell-related article publications in the last two decades, and it indicates the high demand of microfluidic devices for single-cell analysis and applications, evidenced by the increase in the number of scientific publications in each year.…”

mentioning

confidence: 99%