Intracellular Delivery of Nanomaterials via an Inertial Microfluidic Cell Hydroporator

Abstract

The introduction of nanomaterials into cells is an indispensable process for studies ranging from basic biology to clinical applications. To deliver foreign nanomaterials into living cells, traditionally endocytosis, viral and lipid nanocarriers or electroporation are mainly employed; however, they critically suffer from toxicity, inconsistent delivery, and low throughput and are time-consuming and labor-intensive processes. Here, we present a novel inertial microfluidic cell hydroporator capable of delivering a wide range of nanomaterials to various cell types in a single-step without the aid of carriers or external apparatus. The platform inertially focuses cells into the channel center and guides cells to collide at a T-junction. Controlled compression and shear forces generate transient membrane discontinuities that facilitate passive diffusion of external nanomaterials into the cell cytoplasm while maintaining high cell viability. This hydroporation method shows superior delivery efficiency, is high-throughput, and has high controllability; moreover, its extremely simple and low-cost operation provides a powerful and practical strategy in the applications of cellular imaging, biomanufacturing, cell-based therapies, regenerative medicine, and disease diagnosis.

Intracellular Labeling with Extrinsic Probes: Delivery Strategies and Applications

“…Reproduced with permission. [ 149 ] Copyright 2018, American Chemical Society. e) Cells are flown toward a sharp tip by which they become punctured, after which they are released again by reversing the flow.…”

“…[ 148 ] To overcome these limitations, Deng et al created an alternative design, termed the inertial microfluidic cell hydroporator (iMCH), in which shear forces are applied to cells by letting them collide with a sharp tip at a T‐junction (Figures 11b and 12d). [ 149 ] Aided by local enhanced fluid‐shear this induced membrane disruption allowing macromolecules to enter the cells. More than 80% of positive cells were successfully labeled with FD3 with cell viability >90%.…”

A review on nanosystems as an effective approach against infections of <em>Staphylococcus aureus</em>

“…But the studies of nanosystems against S. aureus infections are not complete and we still face the challenges from nanosystems, reasonable largescale production, and so on. There are some contradictions between the absence of enhanced activity against intracellular pathogens and intracellular accumulation of some antibacterial drugs through nanoparticles because of the dormant or quiescent state of bacteria 112,113 and the drug inactivation in the intracellular unfavorable environment. Compared to the free rifampin, the activity against mycobacterium was not enhanced by the polyisobutyl cyanoacrylate nanoparticle, although the amount of intracellular rifampin was increased by the nanoparticles.…”

A Molecular Hero Suit for In Vitro and In Vivo DNA Nanostructures

“…The utilization of the cationic polymer spermidine can protect DNA nanostructures against the denaturing effects of electricity required to electroporate Jurkat cells . An approach that bypasses the classical methods' unfavorable conditions is the use of a microfluidic device . The device focuses cells to a T‐junction, where pores in the cell membrane are generated upon collision with a small protrusion on the device wall, and introduced nanomaterials can then enter the cell.…”