ACS Appl Mater Interfaces. 2014 Dec 10;6(23):20709-16.

Electrically controlled delivery of cargo into single human neural stem cell.

Kim TH, Cho HY, Lee KB, Kim SU, Choi JW.

Department of Chemical & Biomolecular Engineering, Sogang University , 35 Baekbeom-ro, Mapo-gu, Seoul 121-742, Korea.

 

Abstract

Nanoprobe-based techniques have emerged as an efficient tool for the manipulation and analysis of single cells. Here, we report a powerful whole-electrical single-cell manipulation tool that enables rapid and controllable delivery of cargo into single neural stem cells with precision monitoring of the cell penetration process using a conductive nanoprobe. The highly electrically sensitive nanoprobes that were fabricated and the indium tin oxide electrode-integrated cell chip were found to be very effective for monitoring the cell penetration process via current changes that appear as spike-like negative currents. Moreover, the assembly of cargoes onto the nanoprobes was controllable and could reach its maximum load in a very short period of time (<10 min) based on the same electrical system that was used for monitoring cell penetration and without the need for any complex chemical linkers or mediators. Even more remarkably, the cargo assembled on the surface of the nanoprobe was successfully released in a very short period of time (<10 s), regardless of the surrounding intracellular or extracellular environments. The monitoring of cell penetration, assembly of quantum dots (QDs), and release of QDs into the intracellular environment were all accomplished using our whole-electrical system that combined a conductive nanoprobe with cell chip technology. This is a novel technology, which can eliminate complex and time-consuming steps owing to chemical modifications, as well as reduce the time needed for the delivery of cargo into the cell cytosol/nucleus during cell penetration, which is very important for reducing cell damage.

KEYWORDS: electrical assembly and release; human neural stem cell; nanoprobe; quantum dots; single-cell manipulation

PMID: 25204202

 

Supplements:

Since cells are complex living organism, single-cell analysis is extremely important to achieve a variety of information that is hard to obtain from large populations of cells or tissues. Interestingly, recent studies reported that the single cells can be effectively manipulated by using etched silicon tips and the atomic force microscopy or carbon nanotube-modified glass pipets and patch clamp manipulator. These works were proven to be excellent for delivering materials of interest into target cells without giving damages on cells; however, since the delivery of cargo into target cells were completely dependent on the enzymatic activity of cells (i.e., disulfide bond breakage mediated by glutathione), it was relatively hard to control the insertion of cargo into cells in quantitative manner, which is quite important to study the relationship between the materials delivered and the corresponding responses of cells.

By utilizing a new developed tool utilizing insulator-coated conductive nanoprobe and cell chip technology, all the important process including cell penetration, cargo assembly and its release into target cells were successfully conducted based on simple electrical methods. Since any kind of complex chemical reactions, laborious process and expensive instruments are not required, this technology can be highly useful for the development of chip-based rapid single-cell manipulation tool that will enable the delivery of materials of interest (e.g., miRNAs/siRNAs, chemotherapeutic agents, nanoparticles, plasmids, differentiation factors) into target cells (Figure 1).

 

jyl fig1

Figure 1. Schematic diagram representing a system to deliver cargo into single cells based on electrical method

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