Efficient in-droplet cell culture and cytomechanics measurement for assessment of human cellular responses to alcohol.

BACKGROUND

Excessive alcohol consumption poses a significant threat to human health, leading to cellular dehydration, degeneration, and necrosis. Alcohol-induced cellular damage is closely linked to alterations in cellular mechanical properties. However, characterizing these changes following alcohol-related injury remains challenging. Moreover, current research on single-cell mechanics often struggles to culture and measure cells within a controlled microenvironment, leading to complex experimental procedures and imprecise results. (63).

RESULTS

In this study, we developed a novel single cell measurement method that combines cell microculture in alcohol-containing solutions with cytomechanics assessment within microdroplets. This approach integrates key operations, including single-cell encapsulation and culture in droplets, droplet reinjection, and cell deformation analysis within droplets, enabling high-throughput and multi-parameter quantification of single-cell mechanical properties. The use of droplets provides a precisely regulated microculture environment, effectively avoiding channel clogging issues. Additionally, the integration of cytomechanics measurement simplifies the analytical process by eliminating the need for complex techniques within the droplets. Gastric mucosal epithelial cells (GES-1) and human umbilical vein endothelial cells (HUVECs) were selected as models for ethanol-induced injury to validate the proposed technique. The results demonstrate a bidirectional response in cellular deformability following ethanol treatment, with cells becoming stiffer at lower ethanol concentrations and softer at higher concentrations. (136).

SIGNIFICANCE

The integration of droplet microfluidics and cell mechanics offers a powerful platform for investigating the underlying mechanisms of ethanol-induced cellular damage. This approach is also applicable for studying changes in cellular mechanical properties by precisely modulating the microculture environment, providing a reliable tool for drug screening and disease modeling in biochemistry and biomedical engineering. (54).