Abstract
In single cell analysis, transport of foreign substances into a cell is an important technique. In particular, for accurate analysis, a method to transport a small amount (pico-liter order) of substance into the cell without leakage while retaining the cell shape is essential. Because the fusion of the cell and the giant liposome is a closed system to the outside, it may be possible to transport a precise, small amount of substances into the cell. Additionally, there is no possibility that a leaked substance would affect other systems. To develop the liposome-cell transportation system, knowledge about the behavior of substances in the liposome and the cell is important. However, only a few studies have observed the substance transport between a liposome and a cell. Here, we report observation of small amount of substance transport into a single C2C12 cell by using a giant liposome. Substance transport occurred by electrofusion between the cell and the giant liposome containing the substance, which is a closed system. First, to observe the electrofusion and substance transport from the moment of voltage application, we fabricated a microfluidic device equipped with electrodes. We introduced suspensions of cells and liposomes into the microfluidic device and applied alternating current (AC) and direct current (DC) voltages for electrofusion. We observed a small amount (22.4 ± 0.1%, 10.3 ± 0.4% and 9.1 ± 0.1%) of fluorescent substance (Calcein) contained in the liposomes was transported into the cell without leakage outside the cell, and we obtained the diffusion coefficient of Calcein in the cell as 137 ± 18 μm2/s. We anticipate that this system and the knowledge acquired will contribute to future realization of more accurate single cell analysis in a wide range of fields.
Highlights
Cells are the smallest unit of living things
We report on electrofusion and a small amount of substance transport between a cell that retains its shape and a giant liposome; this is a closed system
We successfully carried out an electrofusion and a small amount of substance transport between a C2C12 cell that held its shape and a giant liposome without leakage of liquid
Summary
Cells are the smallest unit of living things. many things are unknown in regard to cells, so that analyses on a single cell scale are very meaningful in the fields of biology and medicine. A functional analysis of genes becomes possible These functional analysis methods are conducted by chemical or electrical treatment of the cell and floating the substances for transport in a dish or micro channel and chamber [4,5,6,7,8,9]. Since it is necessary to suspend a large amount of substances in a volume that is larger than the volume of the cell, they are unsuitable for transport of rare substances In consideration of these problems, a method is required to transport a precise amount of substance on a single cell scale for accurate analysis. The transport observation of this study will be useful for research on liposomes
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