Dielectrophoresis-assisted 3D nanoelectroporation for non-viral cell transfection in adoptive immunotherapy

Chang, Lingqian; Gallego‐Perez, Daniel; Zhao, Xi; Bertani, Paul; Yang, Zhaogang; Chiang, Chi‐Ling; Malkoc, Veysi; Shi, Junfeng; Sen, Chandan K.; O’Donnell, Lynn; Yu, Jianhua; Lu, Wu; Lee, L. James

doi:10.1039/c5lc00553a

Cited by 95 publications

(79 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These scavengers were purchased from Sigma-Aldrich (St. Louis, MO, USA), including sodium pyruvate for H 2 O 2 50; mannitol for OH50; tiron for O23456789101112131415161718192021222324252627282930313233343536373839404142434445464748495051; and N-acetyl cysteine (NAC) as a general ROS scavenger5253. These scavengers are specific with little cross-reactivity to other ROS, so they are used in many protocols for investigating particular ROS functions.…”

Section: Methodsmentioning

confidence: 99%

Intracellular ROS mediates gas plasma-facilitated cellular transfection in 2D and 3D cultures

Wang

et al. 2016

Sci Rep

View full text Add to dashboard Cite

This study reports the potential of cold atmospheric plasma (CAP) as a versatile tool for delivering oligonucleotides into mammalian cells. Compared to lipofection and electroporation methods, plasma transfection showed a better uptake efficiency and less cell death in the transfection of oligonucleotides. We demonstrated that the level of extracellular aqueous reactive oxygen species (ROS) produced by gas plasma is correlated with the uptake efficiency and that this is achieved through an increase of intracellular ROS levels and the resulting increase in cell membrane permeability. This finding was supported by the use of ROS scavengers, which reduced CAP-based uptake efficiency. In addition, we found that cold atmospheric plasma could transfer oligonucleotides such as siRNA and miRNA into cells even in 3D cultures, thus suggesting the potential for unique applications of CAP beyond those provided by standard transfection techniques. Together, our results suggest that cold plasma might provide an efficient technique for the delivery of siRNA and miRNA in 2D and 3D culture models.

show abstract

Section: Methodsmentioning

confidence: 99%

Intracellular ROS mediates gas plasma-facilitated cellular transfection in 2D and 3D cultures

Wang

et al. 2016

Sci Rep

View full text Add to dashboard Cite

show abstract

“…Thus, changes in the method of cell seeding or manipulation cannot significantly improve the transfection efficiency on track-etched membranes. Researchers, including our group, have designed various on-chip manipulation techniques to support 3D EP, including magnetic tweezers (MTs), dielectrophoresis (DEP), and microfluidics 11,13,[24][25][26] . However, instrument-dependent techniques (i.e., MTs and DEP) usually cause irreversible cell damage due to thermal issues, pH changes, and other changes in the microenvironment 27,28 .…”

Section: Introductionmentioning

confidence: 99%

On-chip multiplexed single-cell patterning and controllable intracellular delivery

et al. 2020

Self Cite

View full text Add to dashboard Cite

Conventional electroporation approaches show limitations in the delivery of macromolecules in vitro and in vivo. These limitations include low efficiency, noticeable cell damage and nonuniform delivery of cells. Here, we present a simple 3D electroporation platform that enables massively parallel single-cell manipulation and the intracellular delivery of macromolecules and small molecules. A pyramid pit micropore array chip was fabricated based on a silicon wet-etching method. A controllable vacuum system was adopted to trap a single cell on each micropore. Using this chip, safe single-cell electroporation was performed at low voltage. Cargoes of various sizes ranging from oligonucleotides (molecular beacons, 22 bp) to plasmid DNA (CRISPR-Cas9 expression vectors, >9 kb) were delivered into targeted cells with a significantly higher transfection efficiency than that of multiple benchmark methods (e.g., commercial electroporation devices and Lipofectamine). The delivered dose of the chemotherapeutic drug could be controlled by adjusting the applied voltage. By using CRISPR-Cas9 transfection with this system, the p62 gene and CXCR7 gene were knocked out in tumor cells, which effectively inhibited their cellular activity. Overall, this vacuumassisted micropore array platform provides a simple, efficient, high-throughput intracellular delivery method that may facilitate on-chip cell manipulation, intracellular investigation and cancer therapy.

show abstract

“…www.nature.com/scientificreports www.nature.com/scientificreports/ conclusions Micro-and nanoscale technologies have been used extensively to probe and/or modulate various aspects of cell biology for medical applications [10][11][12][13][14][15][26][27][28][29][30][31][32][33][34][35][36] , especially in cancer therapy and diagnostics 3,37-42 . Here we used microscale engineering tools to demonstrate that tumor-associated MDSCs exhibit structurally guided migration patterns, similar to invasive cancerous cells.…”

Section: Mdsc Subpopulations Exhibit Different Dissemination Capabilimentioning

confidence: 99%

Guided migration analyses at the single-clone level uncover cellular targets of interest in tumor-associated myeloid-derived suppressor cell populations

Duarte‐Sanmiguel

Shukla

Benner

et al. 2020

Sci Rep

Self Cite

View full text Add to dashboard Cite

Myeloid-derived suppressor cells (MDScs) are immune cells that exert immunosuppression within the tumor, protecting cancer cells from the host's immune system and/or exogenous immunotherapies.While current research has been mostly focused in countering MDSC-driven immunosuppression, little is known about the mechanisms by which MDSCs disseminate/infiltrate cancerous tissue. This study looks into the use of microtextured surfaces, coupled with in vitro and in vivo cellular and molecular analysis tools, to videoscopically evaluate the dissemination patterns of MDSCs under structurally guided migration, at the single-cell level. MDSCs exhibited topographically driven migration, showing significant intra-and inter-population differences in motility, with velocities reaching ~40 μm h −1 . Downstream analyses coupled with single-cell migration uncovered the presence of specific MDSC subpopulations with different degrees of tumor-infiltrating and anti-inflammatory capabilities. Granulocytic MDSCs showed a ~≥3-fold increase in maximum dissemination velocities and traveled distances, and a ~10-fold difference in the expression of pro-and anti-inflammatory markers. Prolonged culture also revealed that purified subpopulations of MDSCs exhibit remarkable plasticity, with homogeneous/sorted subpopulations giving rise to heterogenous cultures that represented the entire hierarchy of MDSC phenotypes within 7 days. These studies point towards the granulocytic subtype as a potential cellular target of interest given their superior dissemination ability and enhanced antiinflammatory activity.The tumor microenvironment is highly heterogeneous in nature, with cancerous cells co-habiting with both stromal and immune cells. Such complex cellular interplay plays a central role in modulating tumor progression. Myeloid-derived suppressor cells (MDSCs), in particular, have been known to exert immunosuppressive activity in the tumor niche, which protects cancerous cells from the host immune system and/or different therapeutic modalities 1,2 . While a lot of research has been devoted to developing advanced drugs and drug delivery systems to target cancerous cells 3-5 , and/or blocking MDSC-driven immunosuppression within the tumor niche 6,7 , less is known about the motility mechanisms by which MDSCs disseminate and colonize the tumor in the first place.MDSCs are innate immune cells that are highly expanded in cancer patients 2 . These cells tend to infiltrate tumors and lymphoid tissues, and their levels correlate with increased tumor burden and limited survival in a variety of malignancies 6-9 . MDSCs specifically contribute to the loss of immune effector cell function and reduce the efficacy of immunotherapies. As such, MDSCs have emerged as an attractive therapeutic target in

show abstract

Dielectrophoresis-assisted 3D nanoelectroporation for non-viral cell transfection in adoptive immunotherapy

Cited by 95 publications

References 39 publications

Intracellular ROS mediates gas plasma-facilitated cellular transfection in 2D and 3D cultures

Intracellular ROS mediates gas plasma-facilitated cellular transfection in 2D and 3D cultures

On-chip multiplexed single-cell patterning and controllable intracellular delivery

Guided migration analyses at the single-clone level uncover cellular targets of interest in tumor-associated myeloid-derived suppressor cell populations

Contact Info

Product

Resources

About