Preparing Viable Single Cells from Human Tissue and Tumors for Cytomic Analysis

Area of Science:

• Single-cell biology
• Immunology
• Biotechnology

Background:

• Mass cytometry is a powerful single-cell analysis technique measuring over 35 features per cell.
• Previous applications of mass cytometry have been largely limited to blood and lymphoid tissues due to ease of cell suspension preparation.
• Solid tumors and tissues present challenges for single-cell isolation and analysis.

Purpose of the Study:

• To establish a protocol for single-cell analysis of solid tumors and tissues using mass cytometry.
• To enable the characterization of rare cell types and novel cellular populations within solid tissues.
• To provide a foundation for ex vivo studies of cellular responses in solid tumors.

Main Methods:

• Development and validation of protocols for dissociating solid tumors and tissues into single-cell suspensions.
• Optimization of cell fixation and cryopreservation methods for mass cytometry.
• Demonstration of mass cytometry for analyzing protein and phospho-protein expression in solid tissue-derived cells.

Main Results:

• Successful generation of single-cell suspensions from solid tumors and tissues suitable for mass cytometry.
• Demonstrated ability to measure >35 features per cell across millions of cells from solid tissues.
• Established protocols for preserving cellular integrity for downstream analysis and functional studies.

Conclusions:

• This unit provides a critical primer for applying mass cytometry to solid tumors and tissues.
• The established protocols expand the utility of mass cytometry for comprehensive single-cell analysis in diverse biological contexts.
• The methodology supports further research into the cellular heterogeneity and function within solid tumors and tissues.