Construction of 3D Cellular Composites with Stem Cells Derived from Adipose Tissue and Endothelial Cells by Use of Optical Tweezers in a Natural Polymer Solution

Abstract

To better understand the regulation and function of cellular interactions, three-dimensional (3D) assemblies of single cells and subsequent functional analysis are gaining popularity in many research fields. While we have developed strategies to build stable cellular structures using optical tweezers in a minimally invasive state, methods for manipulating a wide range of cell types have yet to be established. To mimic organ-like structures, the construction of 3D cellular assemblies with variety of cell types is essential. Our recent studies have shown that the presence of nonspecific soluble polymers in aqueous solution is the key to creating stable 3D cellular assemblies efficiently. The present study further expands on the construction of 3D single cell assemblies using two different cell types. We have successfully generated 3D cellular assemblies, using GFP-labeled adipose tissue-derived stem cells and endothelial cells by using optical tweezers. Our findings will support the development of future applications to further characterize cellular interactions in tissue regeneration.

Nonspecific characteristics of macromolecules create specific effects in living cells

“…As shown in the above molecular systems, cells can be cooperatively well-organized into ordered assemblies through the use of macromolecular crowding. Yoshikawa and co-workers showed that, in dextran solution, mammalian cells can be firmly deposited with the desired arrangement using optical manipulation (Hashimoto et al 2016;Yoshida et al 2017;Yamazaki et al 2019). Moderate crowding generated by dextran in the exterior provides an opportunity for rearrangement through which cell-cell association in the assembly can be reinforced.…”

Optically driven microtools with an antibody‐immobilised surface for on‐site cell assembly

“…The processing time of 10 s is sufficient for cell assembly; this is a shorter duration than that of the previous studies on cell assembly. [ 27 , 28 ].…”

“…The design flexibility of the microtool potentially allows us to align individual cells into a cell cluster with desired positions and with a specific number of cells. There are several studies that have observed cell‐cell interactions and co‐culture micro‐environments by assembling cells, including those based on polymer solution, [ 28 ] and cell surface treatment. [ 31 ] The technique proposed here has advantages in terms of its assembly speed (i.e., ∼5 s) and selective cell capture and handling by antibody function.…”