High-throughput functional profiling of single adherent cells via hydrogel drop-screen

Area of Science:

• Biomaterials Science
• Cell Biology
• Biotechnology

Background:

• Single-adherent-cell phenotyping on extracellular matrix (ECM) is crucial for understanding cellular functions related to medical treatments and metastasis.
• Existing platforms lack high-throughput screening capabilities for detailed cellular analysis.

Purpose of the Study:

• To develop a novel hydrogel drop-screen device for rapid, large-scale single-cell morphology and secretion analysis.
• To profile cell phenotypes and investigate functional heterogeneities in response to mechanical stimuli.

Main Methods:

• Cells were anchored to microfluidically fabricated gelatin particles for mechanical stimulation.
• Cellular morphology was assessed by quantifying cytoskeleton expression.
• Droplet encapsulation enabled multiplexed secretion analysis of ADAMs and MMPs at high throughput.

Main Results:

• The device achieved a throughput of approximately 102 cells per second for secretion analysis.
• Distinct functional heterogeneities were observed, correlating with extracellular mechanical stimulations.
• Increased substrate stiffness led to higher cell heterogeneity.

Conclusions:

• The developed hydrogel drop-screen device facilitates high-throughput single-cell phenotyping.
• A database of breast cell lines (MCF10A, MCF-7, MDA-MB-231) was constructed, showing correlations between morphology, secretions, and tumor metastasis.
• The findings highlight the impact of mechanical environment on cell behavior and its relevance to cancer progression.