Single Cell Array Enclosed with a Photodegradable Hydrogel in Microwells for Image-Based Cell Classification and Selective Photorelease of Cells

Abstract

Single cell arrays provide an accurate classification of analyte cells through an image-based analysis of cellular phenotypes. Light-guided cell retrieval from a single cell array is a promising approach for the rapid and simple sorting of difficult to distinguish cells. In this study, we developed a single cell array enclosed with a photodegradable hydrogel in microwells to enable both comprehensive image-based single cell analysis and lightguided cell retrieval. In this system, individual cells became trapped in the microwells together with the photodegradable hydrogel at a high cell density on a chip regardless of cell type, adhesiveness, and motility. Fluorescence-stained model cells and vaccinated dendritic cells were identified by microscopic imaging and then selectively released through the light-induced degradation of the cell-embedding hydrogels. The target cells were selectively retrieved with a purity of >95% from the cell mixture through rapid photorelease, and the retrieved cells were confirmed to grow normally. Our results provide proof-of-principle that the photoresponsive microwell array serves as a versatile tool for image-based cell sorting in cellular researches and the manufacturing processes of high-performance cells.

Dynamic DNA Nanostructures for Cell Manipulation

“…For example, Okamoto et al developed a photocleavable nitrophenyl cross-linker that triggered hydrogel formation to trap cells, while UV light degraded the hydrogel and retrieved the embedded cells with a purity over 95%. 132 The hydrogel system was developed based on bio-orthogonal SPAAC reaction (strain-promoted azide− alkyne cycloaddition), which in principle can reduce cell damage in the system using nonbiorthogonal reactions such as the amide reaction.…”

Star polymer networks: a toolbox for cross-linked polymers with controlled structure

“…These features make SPAAC suitable for functional SPNs for biological applications. [158][159][160][161][162][163][164][165][166][167][168][169][170][171][172][173][174] Johnson and coworkers were the first to utilize SPAAC for SPN syntheses. 175 They modified the synthetic scheme in their pioneering work, 8 which we introduced earlier in this section, to use SPAAC in place of CuAAC.…”

“…Yamaguchi and coworkers developed a photodegradable PEG SPN hydrogel system for single cell trapping and light-guided retrieval. 173 4f-PEG-azide and 4f-PEG-DIBAC (DIBAC = dibenzoazacyclooctyne) were synthesized ( M n = 20 kDa). 4f-PEG-azide had a photolabile o -nitrobenzyl moiety between PEG and the azide group.…”

Selective cell retrieval method using light-responsive gas-generating polymer-based microarrays

“…Therefore, it is not always possible to construct the most suitable surface for each cultured cell especially when we use the photodegradable hydrogels. [15][16][17] In contrast, LGP microarrays utilize the physical force of bubbles to release cells, and so there is no restriction on the type of surface coating material. As such, LGP microarrays are more versatile than other methods for cell capture and can be used for a wider range of cell types.…”

“…16 Yamaguchi et al fabricated the single cell array that enclosed photodegradable hydrogel in microwells. 17 These methods have a great potential to retrieve target single-cell efficiently. However, since these technologies require the use of photodegradable hydrogels, it is extremely difficult to create the ideal culture environment suitable for each kind of cultivated cells by coating the specific extracellular matrix components (ECM) protein (e.g., collagen, fibronectin).…”